Cephem compounds

ABSTRACT

An antibacterial compound                    
     wherein                    
     represents benzene ring, pyridine ring, pyrazine ring or 5-membered aromatic neterocycle (having one oxygen or sulfur atom as ring-constituting atom), there being no R 4  where                    
      represents 5-membered aromatic heterocycle; 
     X and Y respectively represent hydrogen atom or CXY represents C═N—OR 5  wherein R 5  represents hydrogen atom, halo C 1 -C 6  alkyl or C 3 -C 7  cycloalkyl; 
     R 1  represents phenyl, furyl, thienyl, thiazolyl (which may be substituted by amino group), tetrazolyl or thiadiazolyl, 
     R 2 , R 3  and R 4  respectively represent hydrogen atom, halogen, hydroxyl group, nitro, C 1 -C 6  alkoxy, trifluoromethyl, isothiuronium C 1 -C 6  alkyl, amino C 1 -C 6  alkyl, halo C 1 -C 6  alkyl, morpholino, piperidino or piperazinyl.

BACKGROUND OF THE INVENTION TECHNICAL FIELD

1. Field of the Invention

The present invention relates to cephem derivatives or pharmaceuticallyacceptable salts thereof useful as antibacterial agents and representedby the formula I:

wherein

represents benzene ring, pyridine ring, pyrazine ring or 5-memberedaromatic heterocycle (having one oxygen or sulfur atom asring-constituting atom);

X and Y respectively represent hydrogen atom or CXY represents C═N—OR₅wherein R₅ represents hydrogen atom, halo C₁-C₆ alkyl or C₃-C₇cycloalkyl;

R₁ represents phenyl, furyl, thienyl, thiazolyl (which may besubstituted with amino group), tetrazolyl or thiadiazolyl,

R₂, R₃ and R₄ respectively represent hydrogen atom, halogen, hydroxylgroup, nitro, C₁-C₆ alkoxy, trifluoromethyl, isothiuronium C₁-C₆ alkyl,amino C₁-C₆ alkyl, halo C₁-C₆ alkyl, morpholino, piperidino orpiperazinyl, there being no R₄ where

 represents 5-membered aromatic heterocycle; or synthetic intermediatesof the compounds of the formula I and represented by the formula II

wherein Z represents a protective group for a carboxyl group and

 R₂, R₃ and R₄ are as defined above.

2. Description of the Background

In recent years, hardly-curable infections due to pathogenic bacteriahaving resistance against antibiotics such as methicillin-resistantStaphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE)have caused serious problems. Vancomycin, which was the only agentagainst which no resistant bacteria had been found, was frequently usedas medicine against MRSA; however, recently, vancomycin-resistantStaphylococcus aureus (MU3) was found.

Among the above-mentioned resistant bacteria, MRSA are typical ashospital infecting bacteria and there are MRSA-carriers in healthyhumans. Usually, these healthy MRSA-carriers are not sick with MRSA;however, they may tend to become sick with MRSA when they have come intocompromised hosts with reduced vital resistance due to operations orother diseases. Once they become sick, a remedy for their sickness isdifficult to effect. Nowadays, arbekacin, which is an aminoglycoside,and vancomycin, which is a glycopeptide, are used for such diseases.However, already, there are resistant bacteria against arbekacin; andresistant bacteria such as the above-mentioned MU3 have been found alsoas to vancomycin only against which there were no resistant bacteria.Such vancomycin-resistant MRSA are found all over Japan so that there isa fear that, in near feature, MRSA infectious diseses will increasewhich cannot be treated with vancomycin.

Although VRE provide world-shaking issues, though they have not beenfound in Japan yet. VRE are bacteria which have resistance againstvarious antibacaterial agents just as MRSA and which derive fromenterococci (E. faecalis and E. faecium) against which in turn onlyvancomycin was effective and which have gained high resistance againstvancomycin, too. It is highly feared that, in the future, VRE willappear in Japan or prevail as imported infections. However, no effectivemedicines are present now and have been developed yet.

Coming of such medical crisis is readily predicted and urgentdevelopment of medicines effective for the resistant bacteria is desiredto avert such crisis.

Cephem-type antibiotics have been proposed as new antibiotics effectivefor MRSA. More specifically, cephem derivatives having cyclicammoniothiovinyl group at the 3-position have been proposed (JapanesePatent Provisional publication (Kokai) Nos. 6-206886 and 7-304779).These compounds are defective both in terms of antibacterial force andtoxicity, perhaps, due to the cyclic ammoniothiovinyl group, thusfailing to provide new medicines.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present, the inventors, who made devoted researches to pursue cephemderivatives effective for MRSA and VRE and not having theabove-mentioned defective ammonio group in structure, have discoveredcephem derivatives of the formula I having at the 3-positionthiopyranylthiovinyl group with condensed aromatic ring and havingexcellent antibacterial activities against MRSA and vancomycin-resistantE. faecalis, thus accomplishing the present invention.

Conventionally known 3-thiovinyl cephem derivatives having antibacterialactivities are, for example, compounds disclosed in the above-mentionedPublications as well as in Japanese Patent Provisional Publication(Kokai) No. 62-17592 and Japanese Patent Publication (Kokoku) No.6-39475. There is no disclosure in the publications on compounds havingat the 3-position thiopyranylthiovinyl group with condensed aromaticring, nor disclosed is the fact that they are effective for MRSA andvancomycin-resistant E. faecalis.

The compounds and synthetic intermediates of the present invention arerespectively represented by the formulae I and II. The terms used fordefinition of letters in these formulae will be defined and exemplifiedin the following.

The term “C₁-C₆” refers to a group having 1 to 6 carbon atoms.

The term “C₃-C₇” refers to a group having 3 to 7 carbon atoms.

The “C₁-C₆ alkyl group” refers to a straight- or branched-chain alkylgroup such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl,n-pentyl or n-hexyl.

The “C₃-C₇ cycloalkyl group” refers to cyclic alkyl group such ascyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cyclopeptyl.

The “C₁-C₆ alkoxy group” refers to a straight- or branched-chain alkoxygroup such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,tert-butoxy, n-pentyloxy or n-hexyloxy.

The “isothiuronium C₁-C₆ alkyl group” refers to the above-mentioned“C₁-C₆ alkyl group” with isothiuronium group coupled to any of thecarbon atoms.

The “amino C₁-C₆ alkyl group” refers to the above-mentioned “C₁-C₆ alkylgroup” with amino group coupled to any of the carbon atoms.

The “halo C₁-C₆ alkyl group” refers to the above-mentioned “C₁-C₆ alkylgroup” with halogen atom connected to any of the carbon atoms.

The “halogen atom” may be fluorine, chlorine, bromine or iodine atom.

The “5-membered aromatic heterocycle” may be aromatic 5-memberedheterocycle having one oxygen or sulfur atom as ring-constituting atomother than carbon atoms such as furan or thiophene.

When

is 5-membered aromatic heterocycle, the thiopyranyl group with condensedring may be a group with any of the following structures.

wherein R₂ and R₃ are as defined above.

The “protective group for the carboxyl group” may be a group which maybe ordinarily utilized in the art and which may be readily removed. Forexample, it may be tri C₁-C₆ alkylsilyl such as trimethylsilyl,benzhydryl, p-methoxybenzyl, tert-butyl, p-nitrobenzyl.

The compounds according to the present invention may be as follows,though the present invention is not limited to these compounds.

7-[2-(2-Thienyl)acetamido]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(7-chloro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-chloro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-chloro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-chloro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(7-methoxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-methoxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-methoxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-methoxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(4-oxo-5-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-5-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-5-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxymino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-5-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(4-oxo-7-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-7-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxytmino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-7-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-7-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thenyl)acetamido]-3-[2-(6,7,8-trifluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(6,7,8-trifluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(6,7,8-trifluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(6,7,8-trifluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(7-hydroxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-hydroxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-hydroxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-hydroxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(7-isothiuroniummethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-(2-Phenylacetamido)-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-isothiuroniummethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl-3-cephem-4-carboxylicacid

7-(2-Phenylacetamido)-3-[2-(7-isothiuroniummethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(6,7-diisothiuroniummethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(6,7-diisothiuroniummethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-(2-Phenylacetamido)-3-[2-(6,7-diisothiuroniummethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(7-morpholino-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-morpholino-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-morpholino-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-morpholino-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(1H-Tetrazol-1-yl)acetamido]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(4-Thiazolyl)acetamido]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxytmino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(4-oxo-7-piperidino-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-7-piperidino-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-7-piperidino-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-7-piperidino-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)aetamido]-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(3-nitro-7-oxo-7H-thieno([3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(6-bromo-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(6-isothiuroniummethyl-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(5-aminomethyl-2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-(2-Phenylacetamido)-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(6-bromo-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-(2-Phenylacetamido)-3-[2-(6-bromo-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxylmino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(3-nitro-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-(2-Phenylacetamido)-3-[2-(3-nitro-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(6-isothiuroniummethyl-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(5-aminomethyl-2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-(2-Phenylacetamido)-3-[2-(5-aminomethyl-2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido)-3-[2-[6-(2-aminoethyl)-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl]thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-[6-(2-aminoethyl)-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl]thiovinyl]-3-cephem-4-carboxylicacid

7-(2-Phenylacetamido)-3-[2-[6-(2-aminoethyl)-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl]thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido)-3-[2-(2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxylmino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido)-3-[2-(7-oxo-7H-thieno[3,4-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thieno[3,4-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thieno[3,4-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thieno[3,4-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[2,3-c]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[2,3-c]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[2,3-c]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[2,3-c]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[2,3-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[2,3-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[2,3-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(7-oxo-7H-thiopyrano[2,3-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(4-oxo-7-(1-piperazinyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-7-(1-piperazinyl)-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-7-(1-piperazinyl)-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-7-(1-piperazinyl)-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(4-oxo-4H-thiopyrano[2,3-b]pyridin-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(4-oxo-4H-thiopyrano[2,3-b]pyridin-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-(2-Phenylacetamido)-3-[2-(4-oxo-4H-thiopyrano[2,3-b]pyridin-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-(2-Thienyl)acetamido]-3-[2-(8-oxo-8H-thiopyrano[2,3-b]pyrazin-6-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-[2-Hydroxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-[2-(8-oxo-8H-thiopyrano[2,3-b]pyrazin-6-yl)thiovinyl]-3-cephem-4-carboxylicacid

7-(2-Phenylacetamido)-3-[2-(8-oxo-8H-thiopyrano[2,3-b]pyrazin-6-yl)thiovinyl]-3-cephem-4-carboxylicacid

The compounds of the formula I, which have vinyl group at the 3-positionof the structure, may include the following cis isomers (i) and transisomers (ii), and the respective isomers and their mixtures may beincluded in the compounds of the invention. The trans isomers arepreferable from the viewpoint of antibacterial force.

When CXY at the 7-position is an imino group of C═N—OR₅ wherein R₅ is asdefined above, the following syn isomers (iii) and anti isomers (iv)exist, the respective isomers and their mixtures being included in thecompounds of the present invention. The syn isomers are preferable fromthe viewpoint of antibacterial force.

wherein R₅ is as defined above.

Moreover, the compounds of the invention may be in the form ofpharmaceutically acceptable salts such as alkali salts, organic ammoniumsalts or acid addition salts. The appropriate alkali salts which can beused include, for example, potassium salt, sodium salt, calcium salt,magnesium salt, barium salt and ammonium salt. The appropriate acidaddition salts which can be used include inorganic salts such ashydrochloride, hydrobromide, sulfate, nitrate and phosphate as well asorganic acid salts such as acetate, oxalate, propionate, glycolate,lactate, pyruvate, malonate, succinate, maleate, fumarate, malate,tartrate, citrate, benzoate, cinnamate, methanesulfonate,benzenesulfonate, p-toluenesulfonate and salicylate.

The synthetic intermediates of the present invention are, as shown bythe formula II, carboxyls which are different from the compounds I ofthe present invention in that they have cephem with protective group atthe 4-position and with amino at the 7-position. However, the latter areidentical with the former in the substituted group at the 3-position andconfiguration of their vinyl groups similarly include cis isomers (i),trans isomers (ii) and their mixtures.

The compounds of the present invention may be prepared by the followingprocedure.

(1) Preparation of the Synthetic Intermediates of the Present Invention

The synthetic intermediates II of the present invention are obtained by,as shown in process (1) of the below-mentioned reaction formula,reacting a cephem compound of the formula III with a mercaptothiopyranderivative of the formula IV in a solvent and, if needed, in thepresence of tertiary amine such as ethyldiisopropylamine, triethylamineor pyridin, separating the obtained reaction product, and removing theprotective group for the amino group according to a usual manner in thepresence of acid such as hydrochloric, formic or p-toluenesulfonic acid.

There is no particular limitation on the solvent employed, providingthat it is not involved with the reaction. For example, tetrahydrofuran,dichloromethane, benzene, ethyl acetate, dimethylformamide, hexamethylphosphoric triamide, dimethyl sulfoxide, acetone or mixture thereof maybe used. Alternatively, the above-mentioned tertiary amine itself may beused as the solvent. The aimed intermediates are obtained by reactingthe 1-5 moles of the compound of the formula IV per mole of the compoundof the formula III at the temperature range of ice cooling to roomtemperature for 1-6 hours.

(2) Preparation of the Comounds of the Present Invention

The compounds I of the present invention are obtained by, as shown inprocess (2) of the below-mentioned reaction formula, reacting thesynthetic intermediate of the formula II according to the presentinvention with a compound of the formula V in a solvent, separating theobtained reaction product, and removing the protective group for thecarboxyl group according to a usual manner in the presence ofhydrochloric acid, aluminium chloride, formic acid, trifluoroacetic acidor p-toluenesulfonic acid. When trifluoroacetic acid is to be used forremoval of the protective group, it is preferably reacted in thepresence of anisole, thioanisole or phenol so as to facilitate thereaction and suppress any side reactions.

There is no particular limitation on the solvent employed, providingthat it is not involved with the reaction. For example, tetrahydrofuran,dichloromethane, chloroform, benzene, ethyl acetate, dimethylformamide,acetone or mixture thereof may be used. The aimed compound is obtainedby reacting 1-3 moles of the compound of the formula V per mole of thecompound of the formula II at the temperature range of ice cooling toroom temperature for 1-6 hours.

[Reaction Scheme]

wherein Boc represents tert-butoxycarbonyl and Tf representstrifluoromethansulfonyl,

R₁, R₂, R₃, R₄, X, Y and Z are as defined above.

Before introduction, the substituted group of the thiopyran compound ofthe formula IV may be reacted with the cephem compound of the formulaIII as needs demand.

The sequence of the processes in the above-mentioned Reaction Scheme maybe varied. For example, the protective group for the amino group isremoved from the compound of the formula III; then, the latter isreacted with the compound of the formula V and further with the compoundof the formula IV. Finally, the protective group for the carboxyl groupis removed from the reaction product, thus producing the compounds ofthe present invention.

Thus obtained compounds of the present invention may be separated andpurified as needs demand, according to an ordinary method such asextraction, condensation, neutralization, filtration, recrystallizationor column chromatography.

Pharamaceutically acceptable salts of the compounds of the presentinvention may be prepared by various methods known in the art concerned.

Cephem compounds of the formula III which are the starting materials inthe above-mentioned processes are known compounds. Mercaptothiopyranderivatives of the formula IV which involve novel materials can bereadily prepared substantially in accordance with the processesdisclosed in Aust. J. Chem. 40, 1179-1190 (1987), Tetrahedron 35,551-556 (1979), Japanese Patent Provisional Publication (Kokai) No.8-73304 or EP 481441.

Next, antibacterial activity of the compounds of the formula I accordingto the present invention will be described. The numbers of testcompounds in this antibacterial test correspond to those in Examplesreferred to hereinafter.

Antibacterial test was performed in substantial accordance with agarplate dilution method based on Standard method of The Japan Society ofChemotherapy to determine minimum inhibitory concentration (MIC) of therespective test compounds against Staphylococcus aureus (S. aureus FDA209P) and vancomycin-resistant enterococci (E. faecalis NCTC-12201).Also determined was MIC₈₀ on clinically separated strains (27 strains)of MRSA. The inoculated amount was 10⁶CFU/ml and flomoxef (FMOX) andvancomycin (VCM) were used as controls.

The results are as shown in Table 1.

TABLE 1 MIC (μg/ml) S. aureus E. faecalis MRSA tested compound FDA 209PNCTC-12201 MIC₈₀ compound 1 0.20 3.13 3.13 compound 2 0.05 0.78 1.56compound 4 0.10 1.56 3.13 compound 5 0.05 1.56 6.25 compound 7 0.0250.78 3.13 compound 8 0.20 12.5  6.25 compound 9 0.025 3.13 3.13 compound10 0.05 6.25 6.25 compound 11 0.10 1.56 3.13 compound 17 0.39 6.25 3.13compound 18 0.39 6.25 3.13 compound 19 0.10 3.13 3.13 compound 20 0.0251.56 0.78 compound 21 0.025 1.56 0.78 compound 22 ≦0.006 6.25 3.13compound 23 0.10 3.13 1.56 compound 24 ≦0.006 3.13 1.56 compound 250.013 0.78 0.78 compound 26 0.025 1.56 1.56 compound 27 0.025 3.13 1.56compound 28 0.0254 6.25 3.13 compound 30 ≦0.006 6.25 3.13 compound 310.10 3.13 1.56 FMOX 0.20 >100     100 VCM 0.78 >1000     1.56

As is clear from the above, the compounds of the present invention ofthe formula I are effective for MRSA and vancomycin-resistant E.faecalis and can be applied for treatment of infections caused by MRSAor other pathogenic bacteria. The compounds of the present invention maybe administered to human or mammal orally or parenterally. In oraladministration, the compounds may be in the form of tablets, coatedtablets, powders, granules, capsules, microcapsules, syrups and thelike; and in parenteral administration, in the form of injections whichmay include soluble freeze-drying form, suppositories and the like. Inthe preparation of these forms, pharmaceutically acceptable excipient,binders, lubricants, disintegrators, suspensions, emulsifiers,antiseptics, isotonics, stabilizers and dispersing agents, for example,lactose, sucrose, starch, dextrin, crystalline cellulose, kaolin,calcium carbonate, talc, magnesium stearate, distilled water,physiological saline solution and amino acid infusion may be used.

The dosage for humans may depend on the condition of the disease to betreated, the age and weight of the patient and the like. A daily dosagefor an adult may be in the range of from 100 to 5,000 mg and may begiven in divided doses 1 to 4 times a day.

BEST MODE FOR CARRYING OUT THE INVENTION

The present invention is more specifically illustrated with reference tothe following preparations and examples. It is to be, however, notedthat the present invention is not limited to these. Unless otherwiserecited, the compounds of the present invention and their syntheticintermediates shown in the preparations and examples have vinyl group atthe 3-position of cephem as trans isomer.

Preparation 1: Preparation of2-mercapto-7-methoxy-4-oxo-4H-1-benzothiopyran

In accordance with the process disclosed in Aust. J. Chem. 40, 1179(1987), the mixture of 2-fluoro-4-methoxyacetophenone (3 g, 22.73 mmol),N,N-dimethylformamide (25 ml), benzene (17.5 ml) and carbon disulfide(3.78 g, 50.01 mmol) was stirred and added with 55% sodium hydride (2.42g, 55.47 mmol) over about 2.5 hours while inner temperature wasmaintained at 15-20° C. After additional stirring for 30 minutes, thereaction mixture was added with methanol (0.5 ml) and further stirredfor 15 minutes. Benzene was removed under heating to increase innertemperature up to 125-130° C.; at that temperature range, the mixturewas heated for 45 minutes and was allowed to cool to room temperature.The reaction mixture was added with water (80 ml) and acetic acid (2.5ml) and extracted with ether (50 ml×2 times); water layer was filteredout and acidified with concentrated hydrochloric acid. The resultingprecipitates were collected by filteration and were recrystallized frommethanol-1,2-dichloroethane (1:1) to obtain 3.8 g (yield: 75%) of thetitled compound.

Melting Point: 199-200° C.

MS m/z: 224(M⁺)

NMR(DMSO-d₆) δ: 3.81(3H, s), 6.97(1H, s), 7.07(1H, dd, J=2.6 Hz, 9.2Hz), 7.20(1H, d, J=2.6 Hz), 8.04(1H, d, J=9.2 Hz)

Starting from the corresponding starting materials, the followingcompounds are obtained in accordance with the procedure of thePreparation 1.

6,7,8-Trifluoro-2-mercapto-4-oxo-4H-1-benzothiopyran

Melting Point: 158-161° C.

MS m/z: 248(M⁺)

NMR(DMSO-d₆) δ: 7.07(1H, s), 8.03-8.12(1H, m)

2-Mercapto-7-(trifluoromethyl)-4-oxo-4H-1-benzothiopyran

Melting Point: 166-169° C.

MS m/z: 262(M⁺)

NMR(CD₃OD) δ: 7.06(1H, s), 7.64(1H, dd, J=1.6 Hz, 8.5 Hz), 7.77(1H, d,J=1.6 Hz), 8.28(1H, d, J=8.5 Hz)

6-Mercapto-8-oxo-8H-thiopyrano[2,3-b]pyrazin

Melting Point: >178° C. (decomp.)

NMR(DMSO-d₆) δ: 7.31(1H, br), 8.81(2H, br)

Preparation 2: Preparation of5-mercapto-7-oxo-7H-thiopyrano-[3,2-b]furan

(1) In accordance with the process disclosed in Tetrahedron, 35, 551(1979), 2-acetyl-3-bromofuran (10.0 g, 53.2 mmol) and carbon disulfide(7.0 g, 93.2 mmol) were dissolved in anhydrous dimethylformamide (70 ml)and the solution was added with 55% sodium hydride (4.0 g, 93.2 mmol)over about 1.5 hours while it was stirred and inner temperature wasmaintained below 10° C. under ice cooling. After further stirring for0.5 hour at the temperature, the solution was added with methanol (1.1ml, 26.6 mmol) and further stirred at room teperature for 40 minutes.Then, the solution was heated and stirred in the boiling water bath for1 hour and allowed to cool to room temperature. Under water cooling, thesolution was added dropwise with methyl iodide (22.6 g, 159.6 mmol) andstirred at room temperature for 24 hours. The solution was injected intoice water, extracted with dichloromethane (300 ml×2 times), washed withwater and dried over anhydrous magnesium sulfate. The solvent wasremoved under reduced pressure and the residue was recrystallized fromethanol using active carbon to obtain 5.9 g (yield: 56%) of5-methylthio-7-oxo-7H-thiopyrano[3,2-b]furan as colorless needles.

Melting Point: 149-150° C.

NMR(CDCl₃) δ: 2.63(3H, s), 6.74(1H, d, J=2.0 Hz), 7.01(1H, s), 7.80(1H,d, J=2.0 Hz)

(2) The obtained compound (5.7 g, 29 mmol) was dissolved indichloromethane and added dropwise with 80% dichloromethane solution(150 ml) of metachloroperbenzoic acid (5.9 g, 27 mmol) under ice-watercooling and over about 45 minutes. After further stirring at thetemperature for one hour, the precipitates were collected by filtration,washed with dichloromethane and dried over air. This solid was stirredfor a while in the aqueous solution (100 ml) of diluted sodiumhydrogencarbonate and collected by filtration, washed with water andacetone, successively, to obtain 3.3 g (yield: 53%) of5-methylsulfinyl-7-oxo-7H-thiopyrano[3,2-b]furan as colorless powder.

Melting Point: 230-232° C.

NMR(DMSO-d₆) δ: 2.99(3H, s), 7.35(1H, d, J=2.0 Hz), 7.46(1H, s),8.40(1H, d, J=2.0 Hz)

(3) The obtained compound (3.3 g, 15 mmol) was dissolved intetrahydrofuran (160 ml), added with about 70% sodium hydrogensulfide(10 g) and stirred at room temperature for 24 hours. The solvent wasremoved under reduced pressure, added with water (150 ml) fordissolution and washed with dichloromethane. The water layer wasacidified with 2N hydrochloric acid under cooling into pH 1-2. Theprecipitates were collected by filtration and washed with water andether, successively, to obtain 2.3 g (yield: 82%) of the titled compoundas yellow-red powder.

Melting Point: >245° C.

NMR(DMSO-d₆) δ: 7.05 and 7.07(2H, m), 8.31(1H, d, J=1.65 Hz)

The following compounds were obtained from the corresponding startingmaterials and in a procedure similar to that of the Preparation 2.

5-Mercapto-7-oxo-7H-thieno[3,2-b]thiopyran

Melting Point: 151-153° C.

NMR(DMSO-d₆) δ: 7.03(1H, s), 7.45(1H, d, J=5.3 Hz), 8.24(1H, d, J=5.3Hz)

2-Chloro-6-mercapto-4-oxo-4H-thieno[2,3-b]thiopyran

Melting Point: >245° C.

NMR(DMSO-d₆) δ: 7.00(1H, s), 7.51(1H, s)

2-Mercapto-4-oxo-4H-thiopyrano[2,3-b]pyridin

Melting Point: 187-190° C. (decomp.)

NMR(DMSO-d₆) δ: 7.13(1H, s), 7.55(1H, dd, J=4.6 Hz, 8.2 Hz), 8.47(1H,dd, J=1.7 Hz, 8.2 Hz), 8.76(1H, dd, J=1.7 Hz, 4.6 Hz)

Preparation 3: Preparation of7-hydroxy-2-mercapto-4-oxo-4H-1-benzothiopyran

(1) 7-Methoxy-2-mercapto-4-oxo-4H-1-benzothiopyran (2.00 g, 8.92 mmol)was dissolved in DMF (15 ml) and gradually added with sodium hydride(505.8 mg, 11.59 mmol) over 30 minutes in the ice bath and undernitrogen atmosphere. After removal of the ice bath and stir for 12hours, the solution was added with methyl iodide (0.83 ml, 13.37 mmol)and further stirred for 3 hours. The reaction liquid was injected intothe cooled aqueous solution (100 ml) of saturated ammonium chloride. Theprecipitated crude crystals were filtrated and fully washed with waterand dried in a desiccator under reduced pressure. The crude crystalswere washed with the ether-hexane solution to obtain 1.77 g (yield: 83%)of 7-methoxy-2-methylthio-4-oxo-4H-1-benzothiopyran as light yellowpowder.

Melting Point: 133-135° C.

NMR(CDCl₃) δ: 2.61(3H, s), 3.89(3H, s), 6.82(1H, s), 6.91(1H, d, J=2.6Hz), 7.65(1H, dd, J=2.6 Hz, 8.9 Hz), 8.38(1H, d, J=8.9 Hz)

(2) The obtained compound (570 mg, 2.39 mmol) was added with hydrobromicacid (20 ml) and refluxed in heating for 28 hours. After allowed tocool, the solution was neutralized with the aqueous solution of 20%sodium hydroxide under cooling in the ice bath. The precipitated crudecrystals were filtrated and fully washed with water and dried in adesiccator under reduced pressur. The crude crystals were washed withether to obtain 520 mg (yield: 97%) of7-hydroxy-2-methylthio-4-oxo-4H-1-benzothiopyran as light purple powder.

Melting Point: 231-232° C.

NMR(DMSO-d₆) δ: 2.69(3H, s), 6.74(1H, s), 7.01(1H, d, J=2.3 Hz),7.05(1H, brs), 8.16(1H, d, J=8.6 Hz), 10.77(1H, s)

(3) The obtained compound (540 mg, 2.40 mmol) and metachloroperbenzoicacid (860 mg, 3.75 mmol) were used to conduct the process similar tothat shown in (2) of the Preparation 2, thus obtaining 559 mg (yield:97%) of 7-hydroxy-2-methylsulfinyl-4-oxo-4H-1-benzothiopyran as yellowpowder.

NMR(DMSO-d₆) δ: 3.08(3H, s), 7.18(1H, dd, J=2.3 Hz, 8.9 Hz), 7.27(1H, d,J=2.3 Hz), 7.33(1H, d, J=2.3 Hz), 8.29(1H, d, J=8.9 Hz), 11.02(1H, brs)

(4) The obtained compound (559 mg, 2.32 mmol) and sodium hydrogensulfide(1.05 g, 13.12 mmol) were used to conduct the process similar to thatshown in (3) of the Preparation 2, thus obtaining 324 mg (yield: 66%) ofthe titled compound as yellow powder.

Melting Point: 160-163° C.

NMR(DMSO-d₆) δ: 7.01-7.07(3H, m), 8.11(1H, d, J=8.6 Hz), 10.91(1H, br)

Preparation 4: Preparation of2-mercapto-7-morpholino-4-oxo-4H-1-benzothiopyran

(1) 7-Fluoro-2-mercapto-4-oxo-4H-1-benzothiopyran(2.12 g, 10.0 mmol),sodium hydride (567.2 mg, 13.0 mmol) and methyl iodide (0.93 ml, 15.0mmol) were used to conduct the process simiar to (1) of the Preparation3, thus obtaining 1.40 g (yield: 62%) of7-fluoro-2-methylthio-4-oxo-4H-1-benzothiopyran as light yellow powder.

Melting Point: 120-121° C.

NMR(CDCl₃) δ: 2.63(3H, s), 6.83(1H, s), 7.18-7.24(2H, m), 8.48(1H, dd,J=5.9 Hz, 9.6 Hz)

(2) The obtained compound (800 mg, 3.53 mmol) was dissolved in pyridine(15 ml), added with morpholine (1.54 ml, 17.67 mmol) and1,8-diazabicyclo[5,4,0]-7-undecene (DBU) (2.57 ml, 17.67 mmol) andrefluxed under heating for 15 hours. The solvent was removed and theresidue was purified by column chromatography [silica gel, methylenechloride-methanol (20:1)] to obtain 815 mg (yield: 78%) of2-methylthio-7-morpholino-4-oxo-4H-1-benzothiopyran as orange powder.

Melting Point: 163-166° C.

NMR(CDCl₃) δ: 2.63(3H, s), 3.32(4H, t, J=4.9 Hz), 3.87(4H, t, J=4.9 Hz),6.76(2H, brs), 7.03(1H, dd, J=2.6 Hz, 9.2 Hz), 8.32(1H, d, J=9.2 Hz)

(3) The obtained compound (815 mg, 2.78 mmol) and metachloroperbenzoicacid (659 mg, 3.05 mmol) were used to conduct the process similar tothat shown in (2) of the Preparation 2, thus obtaining 546 mg (yield:53%) of 2-methylsulfinyl-7-morpholino-4-oxo-4H-1-benzothiopyran asyellow powder.

Melting Point: 198-200° C.

NMR(CDCl₃) δ: 2.94(3H, s), 3.37(4H, t, J=4.9 Hz), 3.88(4H, t, J=4.9 Hz),6.91(1H, d, J=2.6 Hz), 7.10(1H, dd, J=2.3 Hz, 9.2 Hz), 7.17(1H, s),8.37(1H, d, J=9.2 Hz)

(4) The obtained compound (500 mg, 1.62 mmol) and sodium hydrogensulfide(622 mg, 8.08 mmol) were used to conduct the process similar to thatshown in (3) of the Preparation 2, thus obtaining 443 mg (yield: 69%) ofthe titled compound as yellow powder.

Melting Point: >250° C.

NMR(DMSO-d₆) δ: 2.44(4H, t, J=5.3 Hz), 3.66(4H, t, J=5.3 Hz), 6.89(1H,s), 6.99(1H, d, J=2.6 Hz), 7.11(1H, dd, J=2.6 Hz, 9.2 Hz), 7.92(1H, d,J=9.2 Hz)

The following compounds were obtained from the corresponding startingmaterials and in the prcedure similar to that shown in the Preparation4.

2-Mercapto-4-oxo-7-piperidino-4H-1-benzothiopyran

Melting Point: 120-122° C.

NMR(DMSO-d₆) δ: 1.60(6H, brs), 3.44(4H, brs), 6.93(1H, s), 7.02(1H, d,J=2.6 Hz), 7.15(1H, dd, J=2.6 Hz, 9.2 Hz), 7.95(1H, d, J=9.2 Hz)

2-Mercapto-7-(4-benzyloxycarbonylpiperazin-1-yl)-4-oxo-4H-1-benzothiopyran

Melting Point: 128-131° C.

NMR(DMSO-d₆) δ: 3.55(4H, brs), 3.62(4H, brs), 5.20(2H, s), 7.04(1H, s),7.13(1H, d, J=2.3 Hz), 7.24(1H, dd, J=2.3 Hz, 8.9 Hz), 7. 46(5H, brs),8.07(1H, d, J=8.9 Hz)

Preparation 5: Preparation of7-hydroxymethyl-2-mercapto-4-oxo-4H-1-benzothiopyran

(1) 7-Trifluoromethyl-2-methylthio-4-oxo-4H-1-benzothiopyran (4 g, 14.48mmol) was added to concentrated sulfuric acid (60 ml) and stirred underheating at 110-120° C. for 20 hours. The reaction mixture was injectedto ice water and the precipitates were collected by filtration,dissolved in methyl cellosolve (200 ml) under heating and bleached andpurified by active carbon. Then, the solvent was removed under reducedpressure to obtain 1.8 g (yield: 50%) of7-carboxy-2-methylthio-4-oxo-4H-1-benzothiopyran as Chinese yellowpowder.

MS m/z: 252(M⁺)

NMR(DMSO-d₆) δ: 2.72(3H, s), 6.88(1H, s), 8.04(1H, dd, J=1.3 Hz, 8.5Hz), 8.28(1H, d, J=1.3 Hz), 8.36(1H, d, J=8.5 Hz)

(2) 7-Carboxy-2-methylthio-4-oxo-4H-1-benzothiopyran (1.48 g, 5.87 mmol)was suspended in tetrahydrofuran (30 ml), added with triethylamine (1ml, 7.17 mmol) and cooled to −10° C. Then, the reaction mixture wasadded dropwise with ethyl chlorocarbonate (0.6 ml, 5.94 mmol) andstirred at 0-5° C. for 0.5 hour. The reaction mixture was filtered andthe insoluble matter was further washed with tetrahydrofuran (10 ml×2times). The washing liquid was mixed with the previous tetrahydrofuransolution. Sodium borohidride (568 mg, 15 mmol) was suspended in water (8ml) and added dropwise with the previously obtained tetrahydrofuransolution over 45 minutes under ice-water cooling. The reaction mixturewas acidified with 2N hydrochloric acid and extracted by ethyl acetate(300 ml), washed with saturated saline solution and dried over anhydrousmagnesium sulfate. The solvent was removed under reduced pressure andthe residue was washed with ether to obtain 800mg (yield: 57%) ofsubstaintially pure7-hydroxymethyl-2-methylthio-4-oxo-4H-1-benzothiopyran.

NMR(CDCl₃) δ: 2.62(3H, s), 4.81(1H, d, J=5.6 Hz), 6.83(1H, s), 7.41(1H,d, J=8.3 Hz), 7.52(1H, s), 8.37(1H, d, J=8.3 Hz)

(3) The obtained compound (800 mg, 3.36 mmol) and metachloroperbenzoicacid (725 mg, 3.36 mmol) were used to conduct the process similar tothat shown in (2) of the Preparation 2, thus obtaining 660 mg (yield:77.3%) of 7-hydroxymethyl-2-methylsulfinyl-4-oxo-4H-1-benzothiopyran.

NMR(CDCl₃) δ: 2.95(3H, s), 4.87(1H, d, J=4.0 Hz), 7.26(1H, s), 7.53(1H,d, J=8.0 Hz), 7.72(1H, s), 8.47(1H, d, J=8.0 Hz)

(4) The obtained compound (660 mg, 2.6 mmol) and sodium hydrogensulfide(1.6 g, 20.8 mmol) were used to conduct the process similar to thatshown in (3) of the Preparation 2, thus obtaining 420 mg (yield: 56%) ofthe titled compound as yellow powder.

Melting Point: >174° C. (decomp.)

NMR(DMSO-d₆) δ: 4.62(2H, s), 7.12(1H, s), 7.47(1H, d, J=8.3 Hz),7.55(1H, s), 8.14(1H, d, J=8.3 Hz)

Preparation 6: Preparation of5-tert-butoxycarbonylamino-methyl-2-chloro-6-mercapto-4-oxo-4H-thieno-[2,3-b]thiopyran

(1) 2-Chloro-6-methylthio-4-oxo-4H-thieno[2,3-b]thiopyran (6.64 g, 0.027mol), diethylamine hydrochloride (19.65 g, 0.24 mol) andparaformaldehyde (17.16 g) were added to an aqueous solution of 80%acetic acid and refluxed under heating for 30 hours. The solvent wasremoved under reduced pressure. The residue was added with ice water andstirred for a while, and supernatant was removed. The remaining viscousmatter was dissolved in dimethylformamide (150 ml), filtered for removalof the insoluble matter. Then, the filtrate was added with water (40ml), methanol (100 ml) and potassium carbonate (37 g, 0.7 mol) andstirred for 35 minutes. Then, ice water (700 ml) was added and theresulting precipitates were collected by filtration. The precipitateswere recrystallized from ethanol to obtain 3.5 g (yield: 46.5%) of2-chloro-5-hydroxymethyl-6-methylthio-4-oxo-4H-thieno[2,3-b]thiopyran.

MS m/z: 278(M⁺)

(2) The obtained compound (2 g, 7.17 mmol) was dissolved indichloromethane (30 ml), added with triethylamine (2 ml, 14.34 mmol)under ice cooling, and added dropwise with methanesulfonyl chloride(0.89 ml, 11.47 mmol). The cooling bath was removed and the reactionmixture was stirred at room temperature for 25 hours and added withdichloromethane (150 ml). The reaction mixture was washed with water (50ml) and dried over anhydrous magnesium sulfate. The solvent was removedunder reduced pressure and the residue was washed with ether to obtain 2g (yield: 94%) of2-chloro-5-chloromethyl-6-methylthio-4-oxo-4H-thieno[2,3-b]thiopyran.

MS m/z: 296(M⁺)

NMR(CDCl₃) δ: 2.69(3H, s), 4.88(2H, s), 7.54(1H, s)

(3) The obtained compound (2 g, 6.73 mmol) was suspended indimethylformamide (40 ml), added with sodium azide (1.31 g, 20.19 mmol)while stirred at room temperature, and further stirred for 3 hours. Thereaction mixture was added with ice water and the resulting precipitateswere recovered by filtration and washed with water to obtain 1.9 g(yield: 93%) of5-azidomethyl-2-chloro-6-methylthio-4-oxo-4H-thieno[2,3-b]thiopyran.

MS m/z: 303(M⁺)

IR (KBr): 2093 (−N₃) cm⁻¹

NMR (CDCl₃) δ: 2.66(3H, s), 4.62(2H, s), 7.55(1H, s)

(4) The obtained compound (1.48 g, 4.87 mmol) was dissolved intetrahydrofuran (40 ml), added with triphenylphosphine (2 g, 7.78 mmol)and water (0.4 ml), successively, and stirred at room temperature for 24hours. Then, the reaction mixture was added withdi-tert-butyldicarbonate (1.7 g, 7.79 mmol) and further stirred at roomtemperature for 24 hours. Then, the reaction mixture was added withdichloromethane (150 ml), washed with water and dried over anhydrousmagnesium sulfate. The solvent was removed under reduced pressure andthe residue was purified by silica gel column chromatography (Wako Gel™C-200: dichloromethane-ethyl acetate=8:1) to obtain 1.5 g (yield: 82%)of5-tert-butoxycarbonylaminomethyl-2-chloro-6-methylthio-4-oxo-4H-thieno[2,3-b]thiopyran.

MS m/z: 377(M⁺)

(5) The obtained compound (500 mg, 1.32 mmol) and metachloroperbenzoicacid (299 mg, 1.39 mmol) were used to conduct the process similar tothat shown in (2) of the Preparation 2, thus obtaining 450 mg (yield:87%) of5-tert-butoxycarbonylaminomethyl-2-chloro-6-methylsulfinyl-4-oxo-4H-thieno[2,3-b]thiopyran.Then, this compound (400 mg, 1.02 mmol) and sodium hydrogensulfide (163mg, 2.1 mmol) were used to conduct the process similar to that shown in(3) of the Preparation 2, thus obtaining 320 mg (yield: 95%) of thetitled compound as yellow powder.

Melting Point: 128-131° C. (decomp.)

NMR(DMSO-d₆) δ: 1.40(9H, s), 4.48(2H, s), 7.53(1H, s)

Preparation 7: Preparation of sodium salt of5-mercapto-7-oxo-6-(2-tetrahydropyranyl)oxymethyl-7H-thieno[3,2-b]thiopyran

(1) 5-Methylthio-7-oxo-7H-thieno-[3,2-b]thiopyran (1.77 g, 8.24 mmol)and dimethylamine hydrochloride (6.05 g, 74.2 mmol), paraformaldehyde(5.29 g) and potassium carbonate (2.28 g) were used to conduct theprocess similar to that shown in (1) of the Preparation 6, thusobtaining 1.54 g (yield: 77%) of6-hydroxymethyl-5-methylthio-7-oxo-7H-thieno[3,2-b]thiopyran.

NMR(CDCl₃) δ: 2.66(3H, s), 3.76(1H, t, J=7.0 Hz), 4.94(2H, d, J=7.0 Hz),7.25(1H, d, J=5.3 Hz), 7.85(lH, d, J=5.3 Hz)

(2) The obtained compound (2.16 g, 8.84 mmol) was suspended indichloromethane (120 ml), added with 3,4-dihydro-2H-pyran (8 ml, 87.68mmol) and p-toluenesulfonic acid monohydrate (32.8 mg, 0.172 mmol),successively, under ice-cooled stirring and stirred at room temperaturefor 4 hours. Then, the reaction mixture was added with dichloromethane(100 ml), washed with an aqueous solution of saturated sodiumhydrogencarbonate (50 ml×2 times) and saturated saline solution,successively, and dried over anhydrous magnesium sulfate. The solventwas removed under reduced pressure and the residue was purified bycolumn chromatography (Wako Gel™ C-200: dichloromethane-ethylacetate=8:1), to obtain 2.26 g (yield: 78%) of5-methylthio-7-oxo-6-(2-tetrahydropyranyl)oxymethyl-7H-thieno[3,2-b]thiopyran.

NMR(CDCl₃) δ: 1.51-1.84(6H, m), 2.66(3H, s), 3.58-3.64(1H, m),3.98-4.09(1H, m), 4.74(1H, d, J=10.6 Hz), 4.87-4.89(1H, m), 5.05(1H, d,J=10.6 Hz), 7.21(1H, d, J=5.3 Hz), 7.76(1H, d, J=5.3 Hz)

(3) The obtained compound (1.26 g, 3.83 mmol) and metachloroperbenzoicacid (660 mg, 3.82 mmol) were used to conduct the process similar tothat shown in (2) of the Preparation 2, thus obtaining 839 mg (yield:64%) of5-methylsulfinyl-7-oxo-6-(2-tetrahydropyranyl)oxymethyl-7H-thieno[3,2-b]thiopyran.

NMR(CDCl₃) δ: 1.56-1.84(6H, m), 3.07(3H, d, J=7.0 Hz), 3.56-3.66(1H, m),3.87-3.98(1H, m), 4.46(1H, d, J=12.2 Hz), 4.74(1H, d, J=12.2 Hz),4.81-4.84(1H, m), 4.99(1H, d, J=12.2 Hz), 5.31(1H, d, J=12.2 Hz),7.39(1H, d, J=5.3 Hz), 7.89(1H, d, J=5.3 Hz)

(4) The obtained compound (482 mg, 1.4 mmol) was dissolved intetrahydrofuran (15 ml) and added with 1N sodium hydrogensulfide (2.1ml) under ice-water cooling, and stirred at room temperature for 30minutes. The solvent was removed under reduced pressure to obtain thetitled compound. This compound is used in the succeeding reactionwithout further purification.

Preparation 8: Preparation of sodium salt of6-bromo-5-mercapto-7-oxo-7H-thieno[3,2-b]thiopyran

(1) 5-Methylthio-7-oxo-7H-thieno[3,2-b]thiopyran (2.14 g, 10 mmol) wasdissolved in acetic acid (140 ml) and added dropwise with bromine (0.96ml). The reaction mixture was stirred at 50° C. for 16 hours. Theresulting precipitates were collected by filtration, washed with waterand dried. The reaction product was purified by columnchromatography(Wako Gel™ C-200: dichloromethane-ethyl acetate=15:1) toobtain 1.98 g (yield: 67%) of6-bromo-5-methylthio-7-oxo-7H-thieno[3,2-b]thiopyran.

NMR(CDCl₃) δ: 2.68(3H, s), 7.26(1H, d, J=5.3 Hz), 7.79(1H, d, J=5.3 Hz).

(2) The obtained compound (2.77 g, 9.4 mmol) and metachloroperbenzoicacid (1.96 g, 9.1 mmol) were used to conduct the procedure similar tothat shown in (2) of the Preparation 2, thus obtaining 2.31 g (yield:79%) of 6-bromo-5-methylsulfinyl-7-oxo-7H-thieno[3,2-b]thiopyran.

NMR(CDCl₃) δ: 3.07(1H, s), 7.44(1H, d, J=5.3 Hz), 7.92(1H, d, J=5.3 Hz)

(3) The obtained compound (430 mg, 1.4 mmol) and 1N sodiumhydrogensulfide (2.1 ml) were used to conduct the procedure similar tothat shown in (4) of the Preparation 7, thus obtaining the titledcompound.

Preparation 9: Preparation of sodium salt of5-mercapto-3-nitro-7-oxo-7H-thieno[3,2-b]thiopyran

(1) Mixed acid of nitric acid (1.51 ml) with sulfric acid (5.15 ml) wascooled to −15° C. and added dropwise with a solution of5-methylthio-7-oxo-7H-thieno[3,2-b]thiopyran (1.07 g, 5 mmol) in sulfricacid (4.9 ml). After further stirring at the temperature for one hourand at room temperature for 5 hours, the reaction mixture was injectedinto ice water (300 ml) and stirred for a while. Then, the resultingprecipitates were collected by filtration, washed with water and dried.The reaction product was washed with diisopropylether to obtain 1.15 g(yield: 83%) of5-methylsulfinyl-3-nitro-7-oxo-7H-thieno[3,2-b]thiopyran.

MS m/z: 275(M⁺)

NMR(DMSO-d₆) δ: 3.06(3H, s), 7.48(1H, s), 9.54(1H, s)

(2) The obtained compound (276 mg, 1 mmol) and 1N sodium hydrogensulfide(1.5 ml) were used to conduct the procedure similar to that shown in (4)of the Preparation 7, thus obtaining the titled compound.

EXAMPLE 1 Preparation of7β-amino-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

(1)7β-Tert-butoxycarbonylamino-3-(2-trifluoromethanesulfonyloxyvinyl)-3-cephem-4-carboxylicacid benzhydryl ester (trans isomer) (3.9 g, 6.1 mmol) was dissolved inanhydrous DMF (80 ml), added with 2-mercapto-4-oxo-4H-1-benzothiopyran(3.5 g, 18.3 mmol) and N-ethyldiisopropylamine (0.63 g, 4.87 mmol),successively, and stirred at room temperature under argon gas atmospherefor 24 hours. The reaction product was added with ethyl acetate (350ml), washed with an aqueous solution of saturated sodiumhydrogencarbonate (150 ml×2 times) and dried over anhydrous magnesiumsulfate. The solvent was removed under reduced pressure and the residuewas purified by column chromatography (Wago Gel™ C-200, methylenechloride-ethyl acetate (8:1)) to obtain 3.56 g (yield: 85.4%) of7β-tert-butoxycarbonylamino-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester as yellow-red froth.

NMR(CDCl₃) δ: 1.47(9H, s), 3.60(1H, d, J=17.8 Hz), 3.70(1H, d, J=17.8Hz), 5.03(1H, d, J=5.0 Hz), 5.30(1H, d, J=9.6 Hz), 5.69(1H, dd, J=5.0Hz, 9.6 Hz), 6.67(1H, d, J=15.5 Hz), 7.00(2H, s), 7.30-7.65(14H, m),8.48(1H, d, J=7.9 Hz)

(2) The obtained compound (3.56 g, 5.2 mmol) was suspended inacetonitrile (100 ml), added with p-toluenesulfonic acid monohydrate (5g, 26.0 mmol) and stirred at room temperature for 2.5 hours. Thereaction mixture was added with ethyl acetate (200 ml) and an aqueoussolution of saturated sodium hydrogencarbonate (100 ml) and stirred fora while. Then, ethyl acetate layer was collected, washed with saturatedsaline solution and dried over anhydrous magnesium sulfate. The solventwas removed under reduced pressure and the residue was crystallized fromether to obtain 1.5 g (yield: 49%) of the titled compound as yellowpowder.

NMR(CDCl₃) δ: 3.60(1H, d, J=17.5 Hz), 3.69(1H, d, J=17.5 Hz), 4.82(1H,d, J=5.3 Hz), 5.01(1H, d, J=5.3 Hz), 6.62(1H, d, J=15.5 Hz), 6.97(1H,s), 7.02(1H, s), 7.22-7.70(14H, m), 8.47(1H, d, J=7.9 Hz)

The following compounds were obtained from the corresponding startingmaterials and in the procedure similar to that shown in the Example 1.

7β-Amino-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.63(2H, ABq, J=17.5 Hz), 4.82(1H, d, J=5.3 Hz), 5.00(1H,d, J=4.9 Hz), 6.59(1H, d, J=15.5 Hz), 6.94(1H, s), 7.01(1H, s),7.19-7.49(13H, m), 8.48(1H, dd, J=5.9 Hz, 8.6 Hz)

7β-Amino-3-[2-(7-chloro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.63(2H, ABq, J=17.8 Hz), 4.82(1H, brs), 5.01(1H, d, J=5.3Hz), 6.68(1H, d, J=15.5 Hz), 6.94(1H, s), 7.02(1H, s), 7.28-7.46(13H,m), 8.39(1H, d, J=8.5 Hz)

7β-Amino-3-[2-(7-methoxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.77(2H, ABq, J=17.5 Hz), 3.90(3H, s), 4.99(1H, d, J=4.9Hz), 5.17(1H, d, J=4.9 Hz), 6.74(1H, d, J=15.5 Hz), 6.95(1H, s),6.99(1H, s), 7.07-7.64(13H, m), 8.40(1H, d, J=8.9 Hz)

7β-Amino-3-[2-(4-oxo-5-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.65(2H, ABq, J=17.5 Hz), 4.99(1H, d, J=4.9 Hz), 5.17(1H,d, J=4.9 Hz), 6.69(1H, d, J=15.5 Hz), 6.98(1H, s), 6.99(1H, s),7.21-7.40(11H, m), 7.64-7.69(2H, m), 7.92(1H, d, J=7.9 Hz)

7β-Amino-3-[2-(4-oxo-7-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.64(2H, ABq, J=17.5 Hz), 4.82(1H, d, J=4.9 Hz), 5.01(1H,d, J=4.9 Hz), 6.58(1H, d, J=15.5 Hz), 7.00(1H, s), 7.03(1H, s),7.28-7.46(11H, m), 7.73-7.79(2H, m), 8.58(1H, d, J=8.2 Hz)

7β-Amino-3-[2-(6,7,8-trifluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.64(2H, ABq, J=17.5 Hz), 4.82(1H, d, J=5.2 Hz), 5.02(1H,d, J=5.2 Hz), 6.56(1H, d, J=15.5 Hz), 6.93(1H, s), 7.02(1H, s),7.26-7.45(11H, m), 8.07-8.15(1H, m)

7β-Amino-3-[2-(7-hydroxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(DMSO-d₆) δ: 3.58(2H, ABq, J=17.5 Hz), 4.83(1H, d, J=4.9 Hz),5.03(1H, d, J=4.9 Hz), 7.00(1H, s), 7.02(1H, s), 7.06(1H, d, J=15.5 Hz),7.12-7.83(13H, m), 8.09(1H, d, J=8.6 Hz), 10.76(1H, brs)

7β-Amino-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.60(2H, ABq, J=17.8 Hz), 4.81(1H, d, J=5.0 Hz), 5.00(1H,d, J=5.0 Hz), 6.55(1H, d, J=15.2 Hz), 6.74(1H, d, J=2.0 Hz), 7.01(1H,s), 7.13(1H, s), 7.20-7.50(11H, m), 7.82(1H, d, J=2.0 Hz)

7β-Amino-3-[2-(7-morpholino-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.28(4H, t, J=4.9 Hz), 3.63(2H, q, J=17.5 Hz), 3.86(4H, t,J=4.9 Hz), 4.80(1H, d, J=5.3 Hz), 4.99(1H, d, J=5.3 Hz), 6.63(1H, d,J=15.5 Hz), 6.77(1H, d, J=2.6 Hz), 7.01(1H, s), 7.04(1H, dd, J=2.3 Hz,8.9 Hz), 7.27(1H, d, J=15.5 Hz), 7.30-7.46(11H, m), 8.32(1H, d, J=8.9Hz)

7β-Amino-3-[2-(4-oxo-7-piperidino-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 1.68(6H, brs), 3.38(4H, brs), 3.63(2H, q, J=17.5 Hz),4.80(1H, d, J=5.3 Hz), 4.90(1H, d, J=5.3 Hz), 6.63(1H, d, J=15.5 Hz),6.75(1H, d, J=2.3 Hz), 6.68(1H, s), 7.01(1H, s), 7.03(1H, dd, J=2.3 Hz,8.9 Hz), 7.21(1H, d, J=15.5 Hz), 7.26-7.39(10H, m), 8.27(1H, d, J=8.9Hz)

7β-Amino-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.58(1H, d, J=17.8 Hz), 3.67(1H, d, J=17.8 Hz), 4.81(1H,d, J=5.3 Hz), 5.00(1H, d, J=5.3 Hz), 6.58(1H, d, J=15.5 Hz), 7.00(1H,s), 7.01(1H, s), 7.24(1H, d, J=5.3 Hz), 7.27-7.46(11H, m), 7.83(1H, d,J=5.3 Hz)

7β-Amino-3-[2-(2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.60(2H, ABq, J=17.8 Hz), 4.81(1H, d, J=5.0 Hz), 4.99(1H,d, J=5.0 Hz), 6.52(1H, d, J=15.2 Hz), 6.98(1H, s), 7.00(1H, s),7.20-7.47(11H, m), 7.64(1H, s)

7β-Amino-3-[2-(4-oxo-7-(4-benzyloxycarbonylpiperazin-1-yl)-4H-1-benzothiopyran-2-yl)-thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.38(4H, brs), 3.60-3.69(6H, m), 4.79(1H, d, J=4.9 Hz),4.98(1H, d, J=4.9 Hz), 5.17(2H, s), 6.61(1H, d, J=15.5 Hz), 6.76(1H, d,J=2.3 Hz), 6.87(1H, s), 7.01(1H, s), 7.02(1H, dd, J=2.3 Hz, 8.9 Hz),7.13-7.46(16H, m), 8.31(1H, dd, J=8.9 Hz)

7β-Amino-3-[2-(8-oxo-8H-thiopyrano[2,3-b]pyrazin-6-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.65(2H, ABq, J=17.8 Hz), 4.83(1H, d, J=5.3 Hz), 5.02(1H,d, J=5.3 Hz), 6.56(1H, d, J=15.2 Hz), 7.11(1H, s), 7.26(1H, s),7.20-7.45(11H, m), 8.70(1H, d, J=2.0 Hz), 8.88(1H, d, J=2.0 Hz)

7β-Amino-3-[2-(6-bromo-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.63(1H, d, J=17.5 Hz), 3.72(1H, d, J=17.5 Hz), 4.85(1H,br), 5.03(1H, d, J=5.3 Hz), 6.56(1H, d, J=15.2 Hz), 7.06(1H, s),7.23-7.43(12H, m), 7.79(1H, d, J=5.3 Hz)

7β-Amino-3-[2-(3-nitro-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

NMR(CDCl₃) δ: 3.60(1H, d, J=17.8 Hz), 3.68(1H, d, J=17.8 Hz), 4.83(1H,d, J=5.0 Hz), 5.02(1H, d, J=5.0 Hz), 6.57(1H, d, J=15.5 Hz), 7.02(2H,s), 7.27-7.46(11H, m), 8.83(1H, s)

EXAMPLE 2 Preparation of7β-amino-3-[2-(7-chloromethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

7β-Tert-butoxycarbonylamino-3-[2-(7-hydroxymethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (330 mg, 0.46 mmol) was suspended in anhydrousbenzene (10 ml), added dropwise with thionyl chloride (0.55 ml, 7.59mmol) while stirred at room temperature. Then, the reaction mixture wasstirred at 50 minutes, added with ice water and extracted with ethylacetate (50 ml). Ethyl acetate layer was washed with saturated salinesolution and dried over magnesium sulfate. The solvent was removed underreduced pressure and the residue was crystallized from isopropyl etherto obtain 300 mg (yield: 89%) of7β-tert-butoxycarbonylamino-3-[2(7-chloromethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. Further, the process similar to that shown in (2)of the Example 1 was conducted to obtain 160 mg (yield: 62%) of thetitled compound.

NMR(CDCl₃): 3.63(2H, ABq, 17.5 Hz), 4.64(2H, s), 4.81(1H, d, J=5.3 Hz),5.01(1H, d, J=5.3 Hz), 6.60(1H, d, J=15.5 Hz), 6.96(1H, s), 7.02(1H, s),7.20-7.60(13H, m), 8.44(1H, d, J=7.9 Hz)

EXAMPLE 3 Preparation of7β-amino-3-[2-(6-chloromethyl-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester

(1)7β-Tert-butoxycarbonylamino-3-[2-[7-oxo-6-(2-tetrahydropyranyl)oxymethyl]-7H-thieno[3,2-b]thiopyran-5-yl]thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (400 mg, 0.5 mmol) was added with an aqueoussolution of 50% acetic acid (80 ml) and stirred at 40° C. for 7 hoursunder heating. The reaction mixture was added with dichloromethane (50ml), washed with water (50 ml×2 times) and dried over anhydrousmagnesium sulfate. The solvent was removed under reduced pressure andthe residue was crystallized by diisopropyl ether to obtain 273 mg(yield: 76%) of7β-tert-butoxycarbonylamino-3-[2-(6-hydroxymethyl-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylic acidbenzhydryl ester.

NMR(CDCl₃) δ: 1.47(9H, s), 3.57(1H, d, J=17.8 Hz), 3.66(1H, d, J=17.8Hz), 4.88(2H, s), 5.01(1H, d, J=5.0 Hz), 5.25(1H, d, J=9.0 Hz),5.64-5.70(1H, m), 6.64(1H, d, J=15.5 Hz), 6.98(1H, s), 7.23(1H, d, J=5.3Hz), 7.29-7.45(11H, m), 7.84(1H, d, J=5.3 Hz)

(2) The obtained compound (273 mg, 0.38 mmol) and thionyl chloride (0.46ml, 6.25 mmol) were used to conduct the procedure similar to that shownin the Example 2, thus obtaining 206.3 mg (yield: 73%) of7β-tert-butoxycarbonylamino-3-[2-(6-chloromethyl-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. Further, the process similar to that shown in (2)of the Example 1 was conducted to obtain 132 mg (yield: 74%) of thetitled compound.

NMR(CDCl₃) δ: 3.58(1H, d, J=17.5 Hz), 3.67(1H, d, J=17.5 Hz), 4.83(1H,d, J=5.0 Hz), 4.88(2H, s), 5.00(1H, d, J=5.0 Hz), 6.59(1H, d, J=15.2Hz), 7.05(1H, s), 7.20(1H, d, J=5.3 Hz), 7.28-7.47(11H, m), 7.82(1H, d,J=5.3 Hz)

EXAMPLE 4 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]3-cephem-4-carboxylicacid (compound 1)

(1)7β-Amino-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (160 mg, 0.27 mmol) was dissolved in anhydrousdichloromethane (5 ml), added with thiophene-2-acetic acid (57.6 mg,0.40 mmol) and dicyclohexylcarbodiimide (82.5 mg, 0.40 mmol) at roomtemperature, successively, and stirred at the temperature for 24 hours.The reaction mixture was added with ethyl acetate (20 ml) and theinsoluble matter was filtered. The filtrate was washed with an aqueoussolution of saturated sodium hydrogencarbonate (20 ml×2 times) andsaturated saline solution, successively, and dried over anhydrousmagnesium sulfate. The solvent was removed under reduced pressure andthe residue was purified by column chromatography (Lobar Column™ Si60,Size A (Merck), dichloromethane-ethyl acetate (8:1)) and evaporated todryness. The resulting crystals were washed with ether to obtain 80 mg(yield: 42%) of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester as light yellow crystals.

NMR(CDCl₃) δ: 3.55(1H, d, J=17.5 Hz), 3.63(1H, d, J=17.5 Hz), 3.87(2H,s),5.03(1H, d, J=5.0 Hz), 5.87(1H, dd, J=5.0 Hz, 9.0 Hz), 6.32(1H, d,J=9.0 Hz), 6.65(1H, d, J=15.5 Hz), 6.97(1H, s), 6.99-7.03(3H, m),7.25-7.65(15H, m), 8.48(1H, d, J=7.9 Hz)

(2) The obtained compound (70 mg, 0.099 mmol) was dissolved indichloromethane (3 ml), added with anisole (0.22 ml, 1.98 mmol) andcooled to 0-5° C. The cooled product was added with trifluoroacetic acid(0.17 ml, 1.98 mmol), stirred at room temperature for 4 hours andre-cooled to 0-5° C. The re-cooled product was added with isopropylether (30 ml) under stirring and the resultant precipitates werecollected by filtration and fully washed with isopropyl ether to obtain48 mg (yield: 89%) of the titled compound as yellow powder.

FAB-MS m/z: 543 [M+H]⁺

NMR(DMSO-d₆) δ: 3.69(1H, d, J=17.5 Hz), 4.05(1H, d, J=17.5 Hz), 5.18(1H,d, J=5.0 Hz), 5.73(1H, dd, J=5.0 Hz, 8.2 Hz), 6.94(2H, m), 7.08(1H, s),7.14(1H, d, J=15.0 Hz), 7.26(1H, d, J=15.0 Hz), 7.35(1H, dd, J=2.0 Hz,4.6 Hz), 7.61(1H, dt, J=1.0 Hz, 8.0 Hz), 7.69(1H, dt, J=1.0 Hz, 8.0 Hz),7.82(1H, dd, J=1.0 Hz, 8.0 Hz), 8.25(1H, dd, J=1.0 Hz, 8.0 Hz), 9.20(1H,d, J=8.2 Hz), 13.80(1H, br)

EXAMPLE 5 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 2)

In the same manner as that in (1) of the Example 4,7β-amino-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (120 mg, 0.20 mmol), thiophene-2-acetic acid (45mg, 0.32 mmol) and dicyclohexylcarbodiimide (66 mg, 0.32 mmol) were usedto obtain 100 mg (yield: 69%) of7-[2-(2-thienyl)acetamide]-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (93 mg, 0.13 mmol), anisole(0.28 ml, 2.56 mmol) and trifluoroacetic acid (0.20 ml, 2.56 mmol) wereused to conduct the procedure similar to that shown in (2) of theExample 4 to obtain 24 mg (yield: 33%) of the titled compound.

FAB-MS m/z: 561 [M+H]⁺

NMR(DMSO-d₆) δ: 3.86(2H, s), 3.95(2H, ABq, J=17.5 Hz), 5.27(1H, d, J=4.9Hz), 5.81(1H, dd, J=4.9 Hz, 8.2 Hz), 7.02-7.06(2H, m), 7.17(1H, s),7.22(1H, d, J=15.5 Hz), 7.33(1H, d, J=15.5 Hz), 7.46(1H, dd, J=1.3 Hz,4.9 Hz), 7.55-7.62(1H, m), 7.98(1H, dd, J=2.3 Hz, 9.4 Hz), 8.44(1H, dd,J=6.0 Hz, 8.9 Hz), 9.28(1H, dd, J=8.2 Hz)

EXAMPLE 6 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(7-chloro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 3)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(7-chloro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (105 mg, 0.20 mmol), thiophene-2-acetic acid (110mg, 0.27 mmol) and dicyclohexylcarbodiimide (55 mg, 0.27 mmol) were usedto obtain 86 mg (yield: 68%) of7-[2-(2-thienyl)acetamide]-3-[2-(7-chloro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (80 mg, 0.11 mmol), anisole(0.23 ml, 2.10 mmol) and trifluoroacetic acid (0.21 ml, 2.70 mmol) wereused to conduct the procedure similar to that shown in (2) of theExample 4, thus obtaining 14mg (yield: 22%) of the titled compound.

FAB-MS m/z: 577 [M+H]⁺

NMR(DMSO-d₆) δ: 3.77(2H, ABq, J=17.9 Hz), 3.85(2H, s), 5.18(1H, d, J=4.9Hz), 5.71(1H, dd, J=4.9 Hz, 8.2 Hz), 6.94-7.03(2H, m), 7.07(1H, d,J=15.2 Hz), 7.09(1H, s), 7.26(1H, d, J=15.2 Hz), 7.37(1H, dd, J=1.3 Hz,4.9 Hz), 7.66(1H, dd, J=2.3 Hz, 8.6 Hz), 8.13(1H, d, J=2.3 Hz), 8.28(1H,d, J=8.6 Hz), 9.18(1H, d, J=8.2 Hz)

EXAMPLE 7 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(7-methoxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 4)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(7-methoxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (117 mg, 0.19 mmol), thiophene-2-acetic acid (41mg, 0.28 mmol) and dicyclohexylcarbodiimide (59 mg, 0.28 mmol) were usedto obtain 107 mg (yield: 76%) of7-[2-(2-thienyl)acetamide]-3-[2-(7-methoxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (90 mg, 0.15 mmol), anisole(0.32 ml, 2.94 mmol) and trifluoroacetic acid (0.23 ml, 2.98 mmol) wereused to conduct the procedure similar to that shown in (2) of theExample 4 to obtain 51 mg (yield: 61%) of the titled compound.

FAB-MS m/z: 573 [M+H]⁺

NMR(DMSO-d₆) δ: 3.77(2H, ABq, J=17.9 Hz), 3.86(2H, s), 3.88(3H, s),5.17(1H, d, J=4.9 Hz), 5.71(1H, dd, J=4.9 Hz, 8.2 Hz), 6.93-6.96(2H, m),6.99(1H, s), 7.08(1H, d, J=15.5 Hz), 7.17(1H, d, J=2.6 Hz, 9.2 Hz),7.22(1H, d, J=15.5 Hz), 7.36(1H, dd, J=1.6 Hz, 4.9 Hz), 7.40(1H, d,J=2.6 Hz), 8.22(1H, d, J=9.2 Hz), 9.16(1 H, d, J=7.9 Hz)

EXAMPLE 8 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-5-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 5)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-5-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (210 mg, 0.32 mmol), thiophene-2-acetic acid (92mg, 0.64 mmol) and dicyclohexylcarbodiimide (132 mg, 0.64 mmol) wereused to obtain 226 mg (yield: 91%) of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-5-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (240 mg, 0.31 mmol),anisole (0.67 ml, 6.10 mmol) and trifluoroacetic acid (0.47 ml, 6.10mmol) were used to conduct the procedure similar to that shown in (2) ofthe Example 4, thus obtaining 89 mg (yield: 47%) of the titled compound.

FAB-MS m/z: 611 [M+H]⁺

NMR(DMSO-d₆) δ: 3.71(2H, s), 3.80(2H, ABq, J=17.8 Hz), 5.12(1H, d, J=4.9Hz), 5.67(1H, dd, J=4.9 Hz, 8.2 Hz), 6.87-6.91(2H, m), 7.04(1H, s),7.05(1H, d, J=15.5 Hz), 7.19(1H, d, J=15.5 Hz), 7.31(1H, dd, J=1.6 Hz,4.6 Hz), 7.87(1H, t, J=2.0 Hz), 7.97(1H, d, J=7.6 Hz), 8.10(1H, d, J=7.6Hz), 9.12(1H, d, J=8.2 Hz)

EXAMPLE 9 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-7-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 6)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-7-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (102 mg, 0.15 mmol), thiophene-2-acetic acid (44mg, 0.31 mmol) and dicyclohexylcarbodiimide (64 mg, 0.31 mmol) were usedto obtain 98 mg (yield: 80%) of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-7-trifluoromethyl-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (90 mg, 0.11 mmol), anisole(0.25 ml, 2.32 mmol) and trifluoroacetic acid (0.18 ml, 2.32 mmol) wereused to conduct the procedure similar to that shown in (2) of theExample 4, thus obtaining 54 mg (yield: 76%) of the titled compound.

FAB-MS m/z: 611 [M+H]⁺

NMR(DMSO-d₆) δ: 3.86(2H, s), 3.96(2H, ABq, J=17.8 Hz), 5.28(1H, d, J=4.9Hz), 5.82(1H, dd, J=4.9 Hz, 8.2 Hz), 7.02-7.07(2H, m), 7.21(1H, d,J=15.2 Hz), 7.25(1H, s), 7.36(1H, d, J=15.5 Hz), 7.45(1H, dd, J=1.6 Hz,4.9 Hz), 8.02(1H, dd, J=2.6 Hz, 8.6 Hz), 8.52(1H, d, J=2.6 Hz), 8.54(1H,d, J=8.6 Hz), 9.27(1H, d, J=7.9 Hz)

EXAMPLE 10 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(6,7,8-trifluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 7)

In the same manner as that shown in (1) the Example 4,7β-amino-3-[2-(6,7,8-trifluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (155 mg, 0.24 mmol), thiophene-2-acetic acid (69mg, 0.48 mmol) and dicyclohexylcarbodiimide (100 mg, 0.48 mmol) wereused to obtain 163 mg (yield: 89%) of7-[2-(2-thienyl)acetamide]-3-[2-(6,7,8-trifluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (152 mg , 0.20 mmol),anisole (0.43 ml, 3.99 mmol) and trifluoroacetic acid (0.31 ml, 3.99mmol) were used to conduct the procedure similar to that shown in (2) ofthe Example 4, thus obtaining 91 mg (yield: 77%) of the titled compound.

FAB-MS m/z: 597 [M+H]⁺

NMR(DMSO-d₆) δ: 3.72(2H, s), 3.93(2H, ABq, J=17.5 Hz), 5.20(1H, d, J=4.9Hz), 5.75(1H, dd, J=4.9 Hz, 8.2 Hz), 6.92-6.99(2H, m), 7.14(1H, d,J=15.5 Hz), 7.28(1H, d, J=15.5 Hz), 7.38(1H, dd, J=1.3 Hz, 6.5 Hz),8.10-8.17(1H, m), 9.19(1H, d, J=8.2 Hz)

EXAMPLE 11 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(7-hydroxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 8)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(7-hydroxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (329 mg, 0.54 mmol), thiophene-2-acetic acid (155mg, 1.09 mmol) and dicyclohexylcarbodiimide (225 mg, 1.09 mmol) wereused to obtain 339 mg (yield: 85%) of7-[2-(2-thienyl)acetamide]-3-[2-(7-hydroxy-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (315 mg, 0.43 mmol),anisole (0.94 ml, 8.69 nmmol) and trifluoroacetic acid (0.67 ml, 8.69mmol) were used to conduct the procedure similar to that shown in (2) ofthe Example 4, thus obtaining 86 mg (yield: 35%) of the titled compound.

FAB-MS m/z: 559 [M+H]⁺

NMR(DMSO-d₆) δ: 3.86(2H, s), 3.99(2H, ABq, J=17.2 Hz), 5.27(1H, d, J=4.9Hz), 5.81(1H, dd, J=4.9 Hz, 8.3 Hz), 7.03(1H, s), 7.03-7.05(2H, m),7.07(1H, dd, J=15.2 Hz), 7.10-7.13(1H, m), 7.33(1H, d, J=15.2 Hz),7.45-7.58(2H, m), 8.44(1H, d, J=8.9 Hz), 9.27(1H, d, J=8.3 Hz),10.90(1H, brs)

EXAMPLE 12 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 9)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (60 mg, 0.104 mmol), thiophene-2-acetic acid (22.3mg, 0.156 mmol) and dicyclohexylcarbodiimide (32.3 mg, 0.156 mmol) wereused to obtain 40 mg (yield: 55%) of7-[2-(2-thienyl)acetamide]-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester as light yellow powder. The obtained compound (39mg, 0.056 mmol), anisole (0.12 ml, 1.12 mmol) and trifluoroacetic acid(0.086 ml, 1.12 mmol) were used to conduct the procedure similar to thatshown in (2) of the Example 4, thus obtaining 23 mg (yield: 77%) of thetitled compound as yellow powder.

FAB-MS m/z: 533 [M+H]⁺

NMR(DMSO-d₆) δ: 3.65(1H, d, J=17.8 Hz), 3.77(2H, s), 3.98(1H, d, J=17.8Hz), 5.16(1H, d, J=5.0 Hz), 5.70(1H, dd, J=5.0 Hz, 8.0 Hz), 6.94(2H, m),7.07(1H, d, J=15.5 Hz), 7.08(1H, d, J=15.5 Hz), 7.19(2H, s+d, J=1.6 Hz),7.34(1H, dd, J=1.6 Hz, 4.6 Hz), 8.27(1H, d, J=1.6 Hz), 9.16(1H, d, J=8.0Hz)

EXAMPLE 13 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(7-morpholino-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 10)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(7-morpholino4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (90 mg, 0.13 mmol), thiophene-2-acetic acid (38mg, 0.27 mmol) and dicyclohexylcarbodiimide (56 mg, 0.27 mmol) were usedto obtain 66 mg (yield: 62%) of7-[2-(2-thienyl)acetamide]-3-[2-(7-morpholino-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (60 mg, 0.08 mmol), anisole(0.17 ml, 1.52 mmol) and trifluoroacetic acid (0.12 ml, 1.52 mmol) wereused to conduct the procedure similar to that shown in (2) of theExample 4, thus obtaining 25 mg (yield: 52%) of the titled compound.

FAB-MS m/z: 628 [M+H]⁺

NMR(DMSO-d₆) δ: 3.42(4H, br), 3.82(4H, br), 3.86(2H, s), 3.97(2H, q,J=17.8 Hz), 5.27(1H, d, J=4.9 Hz), 5.81(1H, dd, J=4.9 Hz, 8.3 Hz),7.01(1H, s), 7.03-7.05(2H, m), 7.18(1H, d, J=15.5 Hz), 7.27-7.33(3H, m),7.46(1H, d, J=4.6 Hz), 8.19(1H, d, J=8.9 Hz), 9.27(1H, d, J=8.3 Hz)

EXAMPLE 14 Preparation of7-[2-hydroxyimino-2-(2-aminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (1.5 g, 2.57 mmol),(Z)-2-trityloxyimino-2-(2-tritylaminothiazol-4-yl)acetate (2.6 g, 3.86mmol) and dicyclohexylcarbodiimide (0.8 g, 3.86 mmol) were used toobtain 2.0 g (yield: 63%) of7-[2-trityloxyimino-2-(2-tritylaminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester as orange powder. The obtained compound (1.8 g,1.45 mmol) was dissolved in a mixed solution of acetic acid (64 ml) andwater (6 ml) and stirred under heating at 40-45° C. for 3.5 hours. Thesolvent was removed under reduced pressure and at the temperature below35° C. and the residue was crystallized by ether to obtain 1.0 g of7-[2-hydroxyimino-2-(2-aminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. Then, the obtained ester was added to a mixedliquid of anisole (4.2 ml, 38.6 mmol) with trifluoroacetic acid (21.2ml, 275.2 mmol) cooled in the ice-water bath and stirred at thattemperature for 30 minutes and further at room temperature for 1.5hours. Then, the reaction mixture was cooled again in the ice-waterbath, added with diisopropyl ether (160 ml) and stirred for 30 minutes.The resulting precipitates were collected by filtration and fully washedwith diisopropyl ether to obtain 834 mg (yield: 97%) of the titledcompound.

FAB-MS m/z: 588 [M+H]⁺

NMR(DMSO-d₆) δ: 3.70(1H, d, J=18.0 Hz), 4.02(1H, d, J=18.0 Hz), 5.25(1H,d, J=4.6 Hz), 5.84(1H, dd, J=4.6 Hz, 8.0 Hz), 6.77(1H, s), 7.08(1H, s),7.13(1H, d, J=15.0 Hz), 7.26(1H, d, J=15.0 Hz), 7.62(1H, t, J=7.0 Hz),7.74(1H, t, J=7.0 Hz), 7.85(1H, d, J=7.0 Hz), 8.30(1H, d, J=7.0 Hz),9.62(1H, d, J=8.0 Hz)

EXAMPLE 15 Preparation of7-[2-(1H-tetrazol-1-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 12)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thioylnyl]-3-cephem-4-carboxylicacid benzhydryl ester (160 mg, 0.27 mmol), 1-tetrazolacetic acid (38.4mg, 0.3 mmol) and dicyclohexylcarbodiimide (61.9 mg, 0.3 mmol) were usedto obtain 30 mg (yield: 16%) of7-[2-(1H-tetrazol-1-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester as orange powder. The obtained compound (30.3 mg,0.044 mmol), anisole (0.095 ml, 0.88 mmol) and trifluoroacetic acid(0.07 ml, 0.88 mmol) were used to conduct the procedure similar to thatshown in (2) of the Example 4, thus obtaining 14.6 mg (yield: 63%) ofthe titled compound.

FAB-MS m/z: 529 [M+H]⁺

NMR(DMSO-d₆) δ: 3.67(1H, d, J=17.8 Hz), 4.03(1H, d, J=17.8 Hz), 5.15(1H,d, J=5.0 Hz), 5.33(2H, s), 5.74(1H, dd, J=5.0 Hz, 8.0 Hz), 7.03(1H, s),7.10(1H, d, J=15.4 Hz), 7.20(1H, d, J=15.4 Hz), 7.57(1H, t, J=7.0 Hz),7.69(1H, t, J=7.0 Hz), 7.81(1H, d, J=7.0 Hz), 8.25(1H, d, J=8.0 Hz),9.31(1H, s), 9.52(1H, d, J=8.0 Hz)

EXAMPLE 16 Preparation of7-[2-(4-thiazolyl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 13)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (160 mg, 0.27 mmol), 2-(4-thiazolyl)acetic acid(60 mg, 0.41 mmol) and dicyclohexylcarbodiimide (85 mg, 0.41 mmol) wereused to obtain 123 mg (yield: 64%) of7-[2-(4-thiazolyl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]3-cephem-4-carboxylicacid benzhydryl ester as orange powder. The obtained compound (118.4 mg,0.17 mmol), anisole (0.37 ml, 3.4 mmol) and trifluoroacetic acid (1.31ml, 17.0 mmol) were used to conduct the procedure similar to that shownin (2) of the Example 4, thus obtaining 75 mg (yield: 81%) of the titledcompound.

FAB-MS m/z: 544 [M+H]⁺

NMR(DMSO-d₆) δ: 3.79(1H, d, J=17.8 Hz), 4.12(1H, d, J=17.8 Hz), 3.87(2H,s), 5.27(1H, d, J=5.0 Hz), 5.85(1H, dd, J=5.0 Hz, 8.0 Hz), 7.18(1H, s),7.23(1H, d, J=15.5 Hz), 7.34(1H, d, J=15.5 Hz), 7.56(1H, d, J=2.0 Hz),7.72(1H, t, J=8.0 Hz), 7.84(1H, t, J=8.0 Hz), 7.96(1H, d, J=7.3 Hz),8.40(1H, d,6 J=8.0 Hz), 9.13(1H, d, J=2.0 Hz), 9.23(1H, d, J=8.0 Hz)

EXAMPLE 17 Preparation of7-[2-(2-aminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 14)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (160 mg, 0.27 mmol),2-(2-tritylaminothiazol-4-yl)acetic acid (168 mg, 0.41 mmol) anddicyclohexylcarbodiimide (85 mg, 0.41 mmol) were used to obtain 180 mg(yield: 69%) of7-[2-(2-tritylaminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester as orange powder. The obtained compound (173 mg,0.18 mmol), anisole (0.39 ml, 3.6 mmol) and trifluoroacetic acid (1.39ml, 18 mmol) were used to conduct the procedure similar to that shown in(2) of the Example 4, thus obtaining 75 mg (yield: 75%) of the titledcompound.

FAB-MS m/z: 559 [M+H]⁺

NMR(DMSO-d₆) δ: 3.66(1H, d, J=18.0 Hz), 4.00(1H, d, J=18.0 Hz), 3.68(2H,s), 5.14(1H, d, J=5.0 Hz), 5.72(1H, dd, J=5.0 Hz, 8.0 Hz), 6.53(1H, s),7.02(1H, s), 7.08(1H, d, J=15.5 Hz), 7.19(1H, d, J=15.5 Hz), 7.57(1H, t,J=8.0 Hz), 7.69(1H, t, J=8.0 Hz), 7.80(1H, d, J=8.0 Hz), 8.25(1H, d,J=8.0 Hz), 9.11(1H, d, J=8.0 Hz),

EXAMPLE 18 Preparation of7-[2-cyclopentyloxyimino-2-(2-aminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 15)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (150 mg, 0.26 mmol),(Z)-2-cyclopentyloxyimino-2-(2-tritylaminothiazol-4-yl)acetic acid (194mg, 0.39 mmol) and dicyclohexylcarbodiimide (80.5 mg, 0.39 mmol) wereused to obtain 180 mg (yield: 65%) of7-[2-cyclopentyloxyimino-2-(2-tritylaminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester as orange powder. The obtained compound (150 mg,0.14 mmol), anisole (0.32 ml, 3.0 mmol) and trifluoroacetic acid (1.6ml, 21 mmol) were used to conduct the procedure similar to that shown in(2) of the Example 4, thus obtaining 90 mg (yield: 98%) of the titledcompound as yellow powder.

FAB-MS m/z: 656 [M+H]⁺

NMR(DMSO-d₆) δ: 1.51-2.00(8H, m), 3.70(1H, d, J=18.0 Hz), 4.05(1H, d,J=18.0 Hz), 4.71(1H, m), 5.25(1H, d, J=4.9 Hz), 5.84(1H, dd, J=4.9 Hz,7.9 Hz), 6.78(1H, s), 7.08(1H, s), 7.15(1H, d, J=15.5 Hz), 7.25(1H, d,J=15.5 Hz), 7.62(1H, t, J=8.0 Hz), 7.74(1H, t, J=8.0 Hz), 7.84(1H, d,J=8.0 Hz), 8.30(1H, d, J=8.9 Hz), 9.63(1H, d, J=7.9 Hz)

EXAMPLE 19 Preparation of7-[2-fluoromethyloxyimino-2-(2-aminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 16)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (150 mg, 0.26 mmol),(Z)-2-fluoromethyloxyimino-2-(2-tritylaminothiazol-4-yl)acetic acid (180mg, 0.39 mmol) and dicyclohexylcarbodiimide (80.5 mg, 0.39 mmol) wereused to obtain 174 mg (yield: 65%) of7-[2-fluoromethyloxyimino-2-(2-tritylaminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester as orange powder. The obtained compound (150 mg,0.15 mmol), anisole (0.32 ml, 3.0 mmol) and trifluoroacetic acid (1.6 g,21 mmol) were used to conduct the procedure similar to that shown in (2)of the Example 4, thus obtaining 80 mg (yield: 86%) of the titledcompound as light brown powder.

FAB-MS m/z: 620 [M+H]⁺

NMR(DMSO-d₆) δ: 3.71(1H, d, J=18.0 Hz), 4.03(1H, d, J=18.0 Hz), 5.26(1H,d, J=5.0 Hz), 5.75 and 5.84(2H, d+dd, J=55 Hz, J=5.0 Hz, 8.0 Hz),6.93(1H, s), 7.07(1H, s), 7.13(1H, d, J=15.0 Hz), 7.26(1H, d, J=15.0Hz), 7.63(1H, td, J=1.5 Hz, 8.0 Hz), 7.74(1H, td, J=1.5 Hz, 8.0 Hz),7.84(1H, d, J=8.0 Hz), 8.31(1H, dd, J=1.5 Hz, 8.0 Hz), 9.87(1H, d, J=8.0Hz)

EXAMPLE 20 Preparation of7-[2-hydroxyimino-2-(2-aminothiazol-4-yl)acetamide]-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 17)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (150 mg, 0.26 mmol),(Z)-2-trityloxyimino-2-(2-tritylaminothiazol-4-yl)acetic acid (263 mg,0.39 mmol) and dicyclohexylcarbodiimide (81 mg, 0.39 mmol) were used toobtain 100 mg (yield: 31.3%) of7-[2-trityloxyimino-2-(2-tritylaminothiazol-4-yl)acetamide]-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester as orange powder. The obtained compound (60 mg,0.049 mmol), anisole (0.1 ml, 0.98 mmol) and trifluoroacetic acid (0.53ml, 6.86 mmol) were used to conduct the procedure similar to that shownin (2) of the Example 4, thus obtaining 18 mg (yield: 64%) of the titledcompound as yellow-brown powder.

FAB-MS m/z: 578 [M+H]⁺

NMR(DMSO-d₆) δ: 3.65(1H, d, J=17.5 Hz), 3.96(1H, d, J=17.5 Hz), 5.24(1H,d, J=5.0 Hz), 5.84(1H, dd, J=5.0 Hz, 8.0 Hz), 6.70(1H, s), 7.10(1H, d,J=15.2 Hz), 7.14-7.40(6H, m), 8.28(1H, brs), 9.55(1H, d, J=8.0 Hz),11.50(1H, brs)

EXAMPLE 21 Preparation of7-[2-hydroxyimino-2-(2-aminothiazol-4-yl)acetamide]-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 18)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (216 mg, 0.36 mmol),(Z)-2-trityloxyimino-2-(2-tritylaminothiazol-4-yl)acetic acid (481 mg,0.72 mmol) and dicyclohexylcarbodiimide (148 mg, 0.72 mmol) were used toobtain 108 mg (yield: 24%) of7-[2-trityloxyimino-2-(2-tritylaminothiazol-4-yl)acetamide]-3-[2-(7-fluoro-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (105 mg, 0.083 mmol),anisole (0.18 ml, 1.67 mmol) and trifluoroacetic acid (0.90 ml, 11.62mmol) were used to conduct the procedure similar to that shown in (2) ofthe Example 4, thus obtaining 37 mg (yield: 74%) of the titled compoundas yellow-brown powder.

FAB-MS m/z: 533 [M+H]⁺

NMR(DMSO-d₆) δ: 3.77(1H, d, J=17.5 Hz), 4.10(1H, d, J=17.5 Hz), 5.33(1H,d, J=4.9 Hz), 5.93(1H, dd, J=4.9 Hz, 7.9 Hz), 6.83(1H, s), 7.17(1H, s),7.34(1H, d, J=15.2 Hz), 7.40(1H, brs), 7.58(1H, dt, J=2.3 Hz, 8.6 Hz),7.98(1H, dd, J=2.6 Hz, 9.2 Hz), 8.46(1H, dd, J=5.9 Hz, 8.1 Hz), 9.69(1H,d, J=7.9 Hz), 11.77(1H, brs)

EXAMPLE 22 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-7-piperidino-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 19)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-7-piperidino-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (115 mg, 0.17 mmol), thiophene-2-acetic acid (49mg, 0.34 mmol) and dicyclohexylcarbodiimide (71 mg, 0.34 mmol) were usedto obtain 90 mg (yield: 66%) of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-7-piperidino-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (80 mg, 0.10 mmol), anisole(0.22 ml, 2.02 mmol) and trifluoroacetic acid (0.15 ml, 2.02 mmol) wereused to conduct the procedure similar to that shown in (2) of theExample 4, thus obtaining 51 mg (yield: 80%) of the titled compound.

FAB-MS m/z: 626 [M+H]⁺

NMR(DMSO-d₆) δ: 1.68(6H, brs), 3.50(4H, brs), 3.85(2H, s), 3.94(2H, ABq,J=17.5 Hz), 5.26(1H, d, J=4.9 Hz), 5.81(1H, dd, J=4.9 Hz, 8.2 Hz),6.97(1H, s), 7.02-7.06(2H, m), 7.18(1H, d, J=15.2 Hz), 7.21(1H, d, J=2.3Hz), 7.26(1H, dd, J=2.3 Hz, 8.9 Hz), 7.29(1H, d, J=15.2 Hz), 7.46(1H,dd, J=1.3 Hz, 5.0 Hz), 8.14(1H, d, J=8.9 Hz), 9.27(1H, d, J=8.2 Hz)

EXAMPLE 23 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 20)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (122 mg, 0.21 mmol), thiophene-2-acetic acid (33mg, 0.23 mmol) and dicyclohexylcarbodiimide (48 mg, 0.23 mmol) were usedto obtain 51 mg (yield: 34%) of7-[2-(2-thienyl)acetamide]-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (49 mg, 0.07 mmol), anisole(0.15 ml, 1.38 mmol) and trifluoroacetic acid (0.30 ml, 4.14 mmol) wereused to conduct the procedure similar to that shown in (2) of theExample 4, thus obtaining 19.4 mg (yield: 51%) of the titled compound.

FAB-MS m/z: 549 [M+H]⁺

NMR(DMSO-d₆) δ: 3.68(1H, d, J=17.8 Hz), 3.77(2H, s), 4.02(1H, d, J=17.8Hz), 5.17(1H, d, J=5.0 Hz), 5.73(1H, dd, J=4.6 Hz, 8.3 Hz),6.94-6.97(2H, m), 7.10(1H, d, J=15.5 Hz), 7.11(1H, s), 7.23(1H, d,J=15.1 Hz), 7.37(1H, dd, J=1.6 Hz, 5.0 Hz), 7.64(1H, d, J=5.3 Hz),8.27(1H, d, J=5.3 Hz), 9.19(1H, d, J=8.3 Hz)

EXAMPLE 24 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 21)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (60 mg, 0.10 mmol), thiophene-2-acetic acid (20.5mg, 0.14 mmol) and dicyclohexylcarbodiimide (29.7 mg, 0.14 mmol) wereused to obtain 30 mg (yield: 42%) of7-[2-(2-thienyl)acetamide]-3-[2-(2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (24 mg, 0.03 mmol), anisole(0.07 ml, 0.64 mmol) and trifluoroacetic acid (0.05 ml, 0.64 mmol) wereused to conduct the procedure similar to that shown in (2) of theExample 4, thus obtaining 13 mg (yield: 70%) of the titled compound.

FAB-MS m/z: 583 [M+H]⁺

NMR(DMSO-d₆) δ: 3.65(1H, d, J=17.8 Hz), 3.77(2H, s), 3.99(1H, d, J=17.8Hz), 5.17(1H, d, J=4.6 Hz), 5.72(1H, dd, J=4.6 Hz, 8.0 Hz), 6.94(2H, m),7.08(1H, d, J=15.2 Hz), 7.10(1H, s), 7.22(1H, d, J=15.2 Hz), 7.35(1H,dd, J=1.7 Hz, 4.6 Hz), 7.57(1H, s), 9.17(1H, d, J=8.0 Hz)

EXAMPLE 25 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-7-(piperazin-1-yl)-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid.hydrochloride (compound 22)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-7-(4-benzyloxycarbonylpiperazin-1-yl)-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (113 mg, 0.14 mmol), thiophene-2-acetic acid (40mg, 0.28 mmol) and dicyclohexylcarbodiimide (58 mg, 0.28 mmol) were usedto obtain 82 mg (yield: 62%) of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-7-(4-benzyloxycarbonylpiperazin-1-yl)-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (80 mg, 0.086 mmol) wasdissolved in anisole (5 ml, 46 mmol), added with anhydrous aluminiumchloride (69 mg, 0.517 mmol) under ice bathing and stirred for 12 hours.The reaction liquid was injected into ice water (15 ml) and acidified by2N hydrochloric acid into pH 1.5. The reaction product was washed withethyl acetate and the water layer was freeze-dried. The residue waspurified by reversed phase column chromatography (Wako Gel™ LP-40C18,Wako Junyaku K.K.) with water-acetonitrile (1:1) to obtain 18 mg (yield:32%) of the titled compound as light yellow powder.

FAB-MS m/z: 627 [M+H—HCl]⁺

NMR(DMSO-d₆) δ: 3.22(4H, brs), 3.24-3.74(6H, m), 3.78(2H, s), 5.11(1H,d, J=5.0 Hz), 5.61(1H, dd, J=5.0 Hz, 8.5 Hz), 6.95(1H, s), 6.97-7.15(2H,m), 7.17-7.26(3H, m), 7.37(1H, d, J=15.2 Hz), 7.38(1H, dd, J=1.3 Hz, 4.9Hz), 8.09(1H, d, J=8.9 Hz), 9.15(1H, d, J=8.5 Hz)

EXAMPLE 26 Preparation of7-[2-hydroxyimino-2-(2-aminothiazol-4-yl)acetamide]-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 23)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (183 mg, 0.31 mmol),(Z)-2-trityloxyimino-2-(2-tritylaminothiazol-4-yl)acetic acid (333 mg,0.5 mmol) and dicyclohexylcarbodiimide (102 mg, 0.5 mmol) were used toobtain 250 mg (yield: 65%) of7-[2-trityloxyimino-2-(2-tritylaminothiazol-4-yl)acetamide]-3-[2-(7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester as yellow powder. The obtained compound (201 mg,0.16 mmol), anisole (0.35 ml, 3.2 mmol) and trifluoroacetic acid (0.74ml, 9.6 mmol) were used to conduct the procedure similar to that shownin (2) of the Example 4, thus obtaining 73.7 mg (yield: 65%) of thetitled compound.

FAB-MS m/z: 707 [M+H]⁺

NMR(DMSO-d₆) δ: 3.68(1H, d, J=17.8 Hz), 3.99(1H, d, J=17.5 Hz), 5.24(1H,d, J=5.0 Hz), 5.85(1H, dd, J=5.0 Hz, 8.0 Hz), 6.75(1H, s), 7.12(1H, d,J=15.2 Hz), 7.23(1H, d, J=15.2 Hz), 7.64(1H, d, J=5.3 Hz), 8.27(1H, d,J=5.3 Hz), 9.61(1H, d, J=8.0 Hz), 11.7(1H,s)

EXAMPLE 27 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(8-oxo-8H-thiopyrano[2,3-b]pyrazin-6-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 24)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(8-oxo-8H-thiopyrano[2,3-b]pyrazin-6-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (105 mg, 0.18 mmol), thiopheneacetic acid(38.2 mg,0.27 mmol) and dicyclohexylcarbodiimide (55.7 mg, 0.27 mmol) were usedto obtain 80 mg (yield: 63%) of7-[2-(2-thienyl)acetamide]-3-[2-(8-oxo-8H-thiopyrano[2,3-b]pyrazin-6-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (65 mg, 0.09 mmol), anisole(0.2 ml, 1.9 mmol) and trifluoroacetic acid (0.15 ml, 1.9 mmol) wereused to conduct the procedure similar to that shown in (2) of theExample 4, thus obtaining 40 mg (yield: 50%) of the titled compound.

FAB-MS m/z: 545 [M+H]⁺

NMR(DMSO-d₆): 3.71(1H, d, J=17.8 Hz), 3.78(2H, s), 4.08(1H, d, J=17.8Hz), 5.19(1H, d, J=5 Hz), 5.74(1H, dd, J=5 Hz, 8.3 Hz), 6.94(2H, m),7.10(1H, d, J=15.5 Hz), 7.21(1H, d, J=15.5 Hz), 7.31(2H, m), 8.88(1H, d,J=2 Hz), 8.95(1H, d, J=2 Hz), 9.20(1H, d, J=8.3 Hz)

EXAMPLE 28 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(6-bromo-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 25)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(6-bromo-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (135.5 mg, 0.2 mmol), thiopheneacetic acid (57.5mg, 0.4 mmol) and dicyclohexylcarbodiimide (62.6 mg, 0.3 mmol) were usedto obtain 71.7 mg (yield: 45%) of7-[2-(2-thienyl)acetamide]-3-[2-(6-bromo-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (45 mg, 0.057 mmol),anisole (0.12 ml, 1.14 mmol) and trifluoroacetic acid (0.09 ml, 1.14mmol) were used to conduct the procedure similar to that shown in (2) ofthe Example 4, thus obtaining 35.5 mg (yield: 99%) of the titledcompound.

FAB-MS m/z: 627 [M+H]⁺

NMR(DMSO-d₆) δ: 3.74(1H, d, J=17.8 Hz), 3.78(2H, s), 4.04(1H, d, J=17.8Hz), 5.22(1H, d, J=5.0 Hz), 5.77(1H, dd, J=5.0 Hz, 8.0 Hz),6.93-6.98(2H, m), 7.07(1H, d, J=15.2 Hz), 7.36-7.38(1H, m), 7.39(1H, d,J=15.2 Hz), 7.68(1H, d, J=5.3 Hz), 8.26(1H, d, J=5.3 Hz), 9.20(1H, d,J=8.0 Hz)

EXAMPLE 29 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(3-nitro-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 26)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(3-nitro-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (125 mg, 0.197 mmol), thiopheneacetic acid(42 mg,0.295 mmol) and dicyclohexylcarbodiimide (61 mg, 0.295 mmol) were usedto obtain 108.8 mg (yield: 73%) of7-[2-(2-thienyl)acetamide]-3-[2-(3-nitro-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (108.8 mg, 0.143 mmol),anisole (0.3 ml, 2.86 mmol) and trifluoroacetic acid (0.7 ml, 8.59 mmol)were used to conduct the procedure similar to that shown in (2) of theExample 4, thus obtaining 50.5 mg (yield: 59%) of the titled compound.

FAB-MS m/z: 594 [M+H]⁺

NMR(DMSO-d₆) δ: 3.68(1H, d, J=17.8 Hz), 3.77(2H, s), 4.04(1H, d, J=17.8Hz), 5.19(1H, d, J=5.0 Hz), 5.73(1H, dd, J=5.0 Hz, 8.0 Hz),6.94-6.96(2H, m), 7.11(1H, d, J=15.5 Hz), 7.22(1H, s), 7.27(1H, d,J=15.5 Hz), 7.34-7.36(1H, m), 9.19(1H, d, J=8.0 Hz), 9.49(1H, s)

EXAMPLE 30 Preparation of7-(2-phenylacetamide)-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 27)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (38 mg, 0.065 mmol), phenylacetic acid (15 mg,0.11 mmol) and dicyclohexylcarbodiimide (23 mg, 0.11 mmol) were used toobtain 42 mg of7-(2-phenylacetamide)-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (40 mg, 0.057 mmol),anisole (0.1 ml, 0.92 mmol) and trifluoroacetic acid (0.1 ml, 1.29 mmol)were used to conduct the procedure similar to that shown in (2) of theExample 4, thus obtaining 15 mg (yield: 50%) of the compound.

FAB-MS m/z: 537 [M+H]⁺

NMR(DMSO-d₆): 3.63(2H, d, J=8.9 Hz), 3.96(2H, ABq, J=17.8 Hz), 5.25(1H,d, J=4.9 Hz), 5.80(1H, dd, J=4.9 Hz, 8.2 Hz), 7.17-7.42(8H, m),7.69-7.75(1H, m), 7.96(1H, d, J=7.9 Hz), 8.40(1H, d, J=7.9 Hz), 9.25(1H,d, J=8.2 Hz)

EXAMPLE 31 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(7-isothiuroniummethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid.formate (compound 28)

(1) In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(7-chloromethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (150 mg, 0.24 mmol), thiopheneacetic acid (50.5mg, 0.36 mmol) and dicyclohexylcarbodiimide (74.3 mg, 0.36 mmol) wereused to obtain 60mg (yield: 33%) of7-[2-(2-thienyl)acetamide]-3-[2-(7-chloromethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (50 mg, 0.066 mmol) wasdissolved in a mixed solvent of ethanol (2.5 ml) with dimethylformamide(1 ml), added with thiourea (75.4 mg, 0.99 mmol) and stirred at roomtemperature for 3 days. The solvent was removed under reduced pressureand the reaction mixture was dissolved in methanol (4 ml). The insolublematter was filtered out and the filtrate was dealt with Cephadex LH-20(charged to have 3 cm in dia. and 7 cm in length: methanol) to collectfirstly flowing yellow fraction. The solvent was removed under reducedpressure and the residue was crystallized with isopropyl ether to obtain44 mg (yield: 80%) of 7-[2-(2-thienyl)acetamide]-3-[2-(7-isothiuroniummethyl-4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester.hydrochloride.

FAB-MS m/z: 797[M+H—HCl]⁺

NMR(DMSO-d₆) δ: 3.71(1H, d, J=18.0 Hz), 3.78(2H, s), 4.10(1H, d, J=18.0Hz), 4.62(2H, s), 5.24(1H, d, J=5.0 Hz), 5.80(1H, dd, J=5.0 Hz, 9.0 Hz),6.90-7.50(17H, m), 7.65(1H, d, J=8.2 Hz), 7.86(1H, s), 8.30(1H, d, J=8.2Hz)

(2) The obtained compound (40 mg, 0.048 mmol) was added to formic acid(0.8 ml), stirred at 40-42° C. for one hour under heating. The solventwas removed under reduced pressure and the resulting product wascrystallized with ether to obtain 30 mg (yield: 99%) of the titledcompound.

FAB-MS m/z: 631[M+H—HCOOH]⁺

NMR(DMSO-d₆+D₂O) δ: 3.68(1H, d, J=17.0 Hz), 3.77(1H, s), 4.04(1H, d,J=17.0 Hz), 4.63(2H, s), 5.18(1H, d, J=5.0 Hz), 5.65(1H, m),6.80-7.50(6H, m), 7.65(1H, d, J=8.5 Hz), 7.88(1H, s), 8.30(1H, d, J=8.5Hz)

EXAMPLE 32 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(6-isothiuroniummethyl-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid.trifluoro acetate (compound 29)

In the same manner as that shown in (1) of the Example 4,7β-amino-3-[2-(6-chloromethyl-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester (132.2 mg, 0.21 mmol), thiopheneacetic acid(44.1mg, 0.31 mmol) and dicyclohexylcarbodiimide (64 mg, 0.31 mmol) were usedto obtain 34 mg (yield: 21%) of7-[2-(2-thienyl)acetamide]-3-[2-(6-chloromethyl-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (34 mg, 0.045 mmol) andthiourea (51 mg, 0.668 mmol) were used to conduct the procedure similarto that shown in (1) of the Example 31, thus obtaining 25.1 mg (yield:67%) of 7-[2-(2-thienyl)acetamide]-3-[2-(6-isothiuroniummethyl-7-oxo-7H-thieno[3,2-b]thiopyran-5-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester.hydrochloride.

NMR(MeOD) δ: 3.71(1H, d, J=17.5 Hz), 3.82(2H, s), 3.90(1H, d, J=17.5Hz), 5.18(1H, d, J=5.0 Hz), 5.79(1H, d, J=5.0 Hz), 6.94-6.96(3H, m),7.22-7.41(13H, m), 7.45(1H, d, J=5.3 Hz), 7.98(2H, s), 8.18(1H, d, J=5.3Hz)

Further, the obtained compound (24.5 mg, 0.029 mmol) anisole (0.06 ml,0.582 mmol) and trifluoroacetic acid (0.1 ml, 1.299 mmol) were used toconduct the procedure similar to that shown in (2) of the Example 4,thus obtaining 12.4 mg (yield: 67%) of the titled compound.

FAB-MS m/z: 637 [M+H—CF₃COOH]⁺

NMR(DMSO-d₆) δ: 3.65(1H, d, J=17.5 Hz), 3.78(2H, s), 3.89(1H, d, J=17.5Hz), 4.56(2H, s), 5.16(1H, d, J=5.0 Hz), 5.69(1H, dd, J=5.0 Hz, 8.0 Hz),6.94-6.96(2H, m), 7.36-7.44(3H, m), 7.68(1H, d, J=5.3 Hz), 8.32(1H, d,J=5.3 Hz), 9.16(1H, d, J=8.0 Hz)

EXAMPLE 33 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-4H-thiopyrano[2,3-b]pyridin-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (compound 30)

In the same manner as that shown in (1) of the Example 1,7-[2-(2-thienyl)acetamide]-3-(2-trifluoromethanesulfonyloxyvinyl-3-cephem-4-carboxylicacid benzhydryl ester (80 mg, 0.12 mmol),2-mercapto-4-oxo-4H-thiopyrano[2,3-b]pyridine (58.7 mg, 0.3 mmol) andN-ethyldiisopropylamine (12.4 mg, 0.096 mmol) were used to obtain7-[2-(2-thienyl)acetamide]-3-[2-(4-oxo-4H-thiopyrano[2,3-b]pyridin-2-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (35 mg, 0.49 mmol), anisole(0.1 ml 0.99 mmol) and trifluoroacetic acid (0.08 ml 0.99 mmol) wereused to conduct the procedure similar to that shown in (2) of theExample 4, thus obtaining 16 mg (yield: 60%) of the titled compound.

FAB-MS m/z: 544 [M+H]⁺

NMR(DMSO-d₆) δ: 3.71(1H, d, J=17.8 Hz), 3.77(2H, s), 4.08(1H, d, J=17.8Hz), 5.19(1H, d, J=5.0 Hz), 5.74(1H, dd, J=5.0 Hz, 8.2 Hz), 6.95(2H, m),7.12(1H, d, J=15.0 Hz), 7.13(1H, s), 7.28(1H, d, J=15.0 Hz), 7.35(1H,dd, J=2.0 Hz, 4.6 Hz), 7.67(1H, dd, J=4.6 Hz, 8.0 Hz), 8.60(1H, dd,J=1.7 Hz, 8.0 Hz), 8.85(1H, dd, J=1.7 Hz, 4.6 Hz), 9.20(1H, d, J=8.2 Hz)

EXAMPLE 34 Preparation of7-[2-(2-thienyl)acetamide]-3-[2-(5-aminomethyl-2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid.trifluoroacetate (compound 31)

In the same manner as that shown in (1) of the Example 1,7-[2-(2-thienyl)acetamide]-3-(2-trifluoromethanesulfonyloxyvinyl)-3-cephem-4-carboxylicacid benzhydryl ester (480 mg, 0.7 mmol),5-tert-butoxycarbonylaminomethyl-2-chloro-4-oxo-6-mercapto-4H-thieno[2,3-b]thiopyran(232 mg, 0.7 mmol) and N-ethyldiisopropylamine (72.4 mg, 0.56 mmol) wereused to obtain 174 mg (yield: 28.3%) of7-[2-(2-thienyl)acetamide]-3-[2-(5-tert-butoxycarbonylaminomethyl-2-chloro-4-oxo-4H-thieno[2,3-b]thiopyran-6-yl)thiovinyl]-3-cephem-4-carboxylicacid benzhydryl ester. The obtained compound (40 mg, 0.046 mmol),anisole (0.1 ml, 0.92 mmol) and trifluoroacetic acid (0.071 ml, 0.92mmol) were used to conduct the procedure similar to that shown in (2) ofthe Example 4, thus obtaining 14 mg (yield: 42%) of the titled compound.

FAB-MS m/z: 612 [M+H—CF₃COOH]⁺

NMR(DMSO-d₆) δ: 3.60(1H, d, J=17.5 Hz), 3.77(2H, s), 3.86(1H, d, J=17.5Hz), 4.11(2H, s), 5.13(1H, d, J=5.0 Hz), 5.68(1H, dd, J=5.0 Hz, 8.0 Hz),6.85(1H, d, J=15.5 Hz), 6.95(2H, m), 7.27(1H, d, J=15.5 Hz), 7.33(11,dd, J=1.5, 4.5 Hz), 7.62(1H, s), 9.14(1H, d)

EXAMPLE 35 Preparation of sodium7-[2-hydroxyimino-2-(2-aminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carbonate(compound 11)

7-[2-hydroxyimino-2-(2-aminothiazol-4-yl)acetamide]-3-[2-(4-oxo-4H-1-benzothiopyran-2-yl)thiovinyl]-3-cephem-4-carboxylicacid (500 mg, 0.85 mmol) was suspended in water (30 ml) and adjusted bysaturated sodium hydrogencarbonate into pH 7.6. The resulting solutionwas separated and purified by reversed phased column chromatography(RP-18, acetonitrile-water (3:7)). The obtained fraction wasfreeze-dried to obtain 260 mg (yield: 50%) of the titled compound.

Melting Point: >300° C. (decomp.)

FAB-MS m/z: 610 [M+Na]⁺

NMR(DMSO-d₆) δ: 3.47(1H, d, J=16.5 Hz), 3.61(1H, d, J=16.5 Hz), 5.05(1H,d, J=5.0 Hz), 5.60(1H, dd, J=5.0 Hz, 9.0 Hz), 6.38(1H, d, J=15.0 Hz),6.60(1H, s), 6.87(1H, s), 7.04(2H, brs), 7.40-7.80(4H, m), 8.23(1H, d,J=8.0 Hz), 9.40(1H, br)

The following compound was obtained by the procedure similar to thatshown in the Example 35.

Sodium7-[2-(2-thienyl)acetamide]-3-[2-(7-oxo-7H-thiopyrano[3,2-b]furan-5-yl)thiovinyl]-3-cephem-4-carbonate

Melting Point: 175-180° C. (decomp.)

FAB-MS m/z: 555 [M+Na]⁺

NMR(DMSO-d₆) δ: 3.50(2H, ABq, J=17.0 Hz), 3.80(2H, s), 5.05(1H, d, J=5.0Hz), 5.55(1H, dd, J=5.0 Hz, 8.0 Hz), 6.45(1H, d, J=15.0 Hz), 6.95(2H,m), 7.05(1H, s), 7.17(1H, d, J=1.6 Hz), 7.35(1H, m), 7.55(1H, d, J=15.0Hz), 8.24(1H, d, J=1.6 Hz), 9.10(1H, d, J=8.0 Hz)

Capability of Exploitation in Industry

The cephem compound according to the present invention has excellentantibacterial activity against MRSA and vancomycin-resistant E. faecalisand therefore is applicable to treatment of MRSA and other infectionsdue to pathogenic bacteria.

What is claimed is:
 1. A cephem compound or pharmaceutically acceptablesalt thereof having the formula (I):

represents a benzene ring, pyridine ring, pyrazine ring, or a 5-memberedaromatic heterocycle having one oxygen or sulfur atom as aring-constituting atom; X and Y each independently represents hydrogenor CXY represents C═N—OR₅, wherein R₅ represents hydrogen, halo C₁-C₆alkyl or C₃-C₇ cycloalkyl; R₁ represents phenyl, furyl, thienyl,thiazolyl which is optionally substituted, tetrazolyl or thiadiazolyl;and R₂, R₃ and R₄ each independently represents hydrogen, halogen,hydroxyl, nitro, C₁-C₆ alkoxy, trifluoromethyl, amino C₁-C₆ alkyl, haloC₁-C₆ alkyl, morpholino, piperidino or piperazinyl; with the provisothat when

 represents 5-membered aromatic heterocycle, no group R₄ is present. 2.The compound of claim 1, wherein

is a benzene ring.
 3. The compound of claim 1, wherein

is a benzene ring; and X and Y each represent hydrogen.
 4. The compoundof claim 1, wherein

is a benzene ring; X and Y are each hydrogen; and R₁ is phenyl, thienylor tetrazolyl.
 5. The compound of claim 1 wherein

is a benzene ring; and CXY is C═N—OR₅, wherein R₅ is hydrogen,fluoromethyl or cyclopentyl.
 6. The compound of claim 1, wherein

is a benzene ring; CXY is C═N—OR₅, wherein R₅ is hydrogen, fluoromethylor cyclopentyl; and R₁ is amino-substituted thiazolyl.
 7. The compoundof claim 1, wherein

is a furan ring.
 8. The compound of claim 1, wherein

is a furan ring; CXY is C═N—OH; and R₁ is amino-substituted thiazolyl.9. The compound of claim 1, wherein

is a thiophene ring.
 10. The compound of claim 1, wherein

is a thiopene ring; X and Y are each hydrogen; and R₁ is thienyl. 11.The compound of claim 1, wherein

is a thiophene ring; CXY is C═N—OH; and R₁ is amino-substitutedthiazolyl.
 12. The compound of claim 1, wherein

is a pyridine ring.
 13. The compound of claim 1, wherein

is a pyridine ring; X and Y are each hydrogen; and R₁ is thienyl. 14.The compound of claim 1, wherein

is a pyrazine ring.
 15. The compound of claim 1, wherein

is a pyrazine ring; X and Y are each hydrogen; and R₁ is thienyl. 16.The compound of claim 1, which is a trans-isomer.
 17. A compound havingthe formula (II):

represents benzene ring, pyridine ring, pyrazine ring, or 5-memberedaromatic heterocycle having one oxygen or sulfur atom as aring-constituting atom; and R₂, R₃ and R₄ each independently representhydrogen, halogen, hydroxyl, nitro, C₁-C₆ alkoxy, trifluoromethyl, haloC₁-C₆ alkyl, morpholino, piperidino or piperazinyl, and Z representsprotective group for carbonyl.
 18. The compound of claim 1, wherein CXYis an imino group of the formula C═N—OR₅, wherein R₅ is as defined,which is a syn isomer having the partial configuration:


19. A pharmaceutical composition, comprising: a) an effective amount ofone or more of the compounds of claim 1; and b) a pharmaceuticallyacceptable carrier.
 20. A method of treating a bacterial infection in amammal, which comprises administering an effective amount of one or morecompounds of claim 1, to a mammal in need thereof.